# US 2019/0316129 A1 · Compositions and Methods for Treating Alpha-1 Antitrypsin Deficiency > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0316129 A1 - Document ID: US20190316129A1 - Status: Published application - Kind: A1 - Application number: 16/446,764 - Filed: 2019-06-20 - Published: 2019-10-17 - Earliest claimed: 2016-12-22 - Assignees: Intellia Therapeutics, Inc. - Applicants: Intellia Therapeutics, Inc. - Inventors: Shobu Odate; Walter Strapps; Reynald Michael Lescarbeau - Web page: https://patentgrep.com/patent/US20190316129A1 ## CPC - C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} - A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) - A61K 9/14: Particulate form, e.g. powders, {Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles (microspheres A61K9/16; microcapsules A61K9/50; nanocapsules, nanoparticles of the matrix type A61K9/51)} - A61K 9/5146: {obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides} - A61K 47/24: containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids - A61K 47/28: Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid - A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics - A61P 11/00: Drugs for disorders of the respiratory system - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C12N 15/86: Viral vectors - C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR] - C12N 2310/315: Phosphorothioates - C12N 2310/344: Position-specific modifications, e.g. on every purine, at the 3'-end - C12N 2310/346: having a combination of backbone and sugar modifications - C12N 2320/32: Special delivery means, e.g. tissue-specific - C12N 2320/51: modulating the chemical stability, e.g. nuclease-resistance - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract Compositions and methods for introducing double-stranded breaks within the SERPINA1 gene are provided. Compositions and methods for reducing and eliminating mutant forms of al-antitrypsin (AAT), such as seen in subjects having al-antitrypsin deficiency (AATD), are provided. ## Claims ### Claim 1 A method of inducing a double-stranded break (DSB) within the SERPINA1 gene, comprising delivering a composition to a cell, wherein the composition comprises a guide RNA comprising a guide sequence selected from SEQ ID NOs: 5-129 or a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID NOs: 5-129. ### Claim 2 A method of modifying the SERPINA1 gene comprising delivering a composition to a cell, wherein the composition comprises (i) an RNA-guided DNA binding agent or a nucleic acid encoding an RNA-guided DNA binding agent and (ii) a guide RNA comprising a guide sequence selected from SEQ ID NOs: 5-129 or a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID NOs: 5-129. ### Claim 3 A method of treating alpha-1 antitrypsin deficiency (AATD), comprising administering a composition to a subject in need thereof, wherein the composition comprises (i) an RNA-guided DNA binding agent or a nucleic acid encoding an RNA-guided DNA binding agent and (ii) a guide RNA comprising a guide sequence selected from SEQ ID NOs: 5-129 or a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID NOs: 5-129, thereby treating AATD. ### Claim 4 A method for reducing or preventing the accumulation of alpha-1 antitrypsin (AAT) in the liver in a subject, comprising administering a composition to a subject in need thereof, wherein the composition comprises (i) an RNA-guided DNA binding agent or a nucleic acid encoding an RNA-guided DNA binding agent and (ii) a guide RNA comprising a guide sequence selected from SEQ ID NOs: 5-129 or a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID NOs: 5-129, thereby reducing accumulation of AAT in the liver. ### Claim 5 A composition comprising a guide RNA comprising a guide sequence selected from SEQ ID NOs: 5-129 or a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID NOs: 5-129. ### Claim 6 A composition comprising a vector encoding a guide RNA, wherein the guide RNA comprises a guide sequence selected from SEQ ID NOs: 5-129 or a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID NOs: 5-129. ### Claim 7 (depends on 5, 6) The composition of claim 5 or 6, for use in inducing a double-stranded break (DSB) within the SERPINA1 gene in a cell or subject. ### Claim 8 (depends on 5, 6) The composition of claim 5 or 6, for use in modifying the SERPINA1 gene in a cell or subject. ### Claim 9 (depends on 5, 6) The composition of claim 5 or 6, for use in treating alpha-1 antitrypsin deficiency (AATD) in a subject. ### Claim 10 (depends on 5, 6) The composition of claim 5 or 6, for use in reducing AAT serum or liver concentration in a subject. ### Claim 11 (depends on 5, 6) The composition of claim 5 or 6, for use in reducing or preventing the accumulation of alpha-1 antitrypsin (AAT) in the liver in a subject. ### Claim 12 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11) The method of any one of claims 1-4 or the composition for use of any one of claims 5-11, wherein the composition reduces serum and/or liver AAT levels. ### Claim 13 (depends on 12) The method or composition for use of claim 12, wherein the serum and/or liver AAT levels are reduced by at least 40% as compared to serum and/or AAT levels before administration of the composition. ### Claim 14 (depends on 12) The method or composition for use of claim 12, wherein the serum and/or liver AAT levels are reduced by 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, 90-95%, 95-98%, 98-99%, or 99-100% as compared to serum and/or AAT levels before administration of the composition. ### Claim 15 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14) The method or composition for use of any one of claim 1-4 or 7-14, wherein the composition results in editing of the SERPINA1 gene. ### Claim 16 (depends on 15) The method or composition for use of claim 15, wherein the editing is calculated as a percentage of the population that is edited (percent editing). ### Claim 17 (depends on 16) The method or composition for use of claim 16, wherein the percent editing is between 30 and 99%. ### Claim 18 (depends on 17) The method or composition for use of claim 17, wherein the percent editing is between 30 and 35%, 35 and 40%, 40 and 45%, 45 and 50%, 50 and 55%, 55 and 60%, 60 and 65%, 65 and 70%, 70 and 75%, 75 and 80%, 80 and 85%, 85 and 90%, 90 and 95%, or 95 and 99%. ### Claim 19 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18) The method or composition for use of any one of claim 1-4 or 7-18, wherein the composition is administered or delivered at least two times. ### Claim 20 (depends on 19) The method or composition for use of claim 19, wherein the composition is administered or delivered at least three times. ### Claim 21 (depends on 19) The method or composition for use of claim 19, wherein the composition is administered or delivered at least four times. ### Claim 22 (depends on 19) The method or composition for use of claim 19, wherein the composition is administered or delivered up to five, six, seven, eight, nine, or ten times. ### Claim 23 (depends on 19, 20, 21, 22) The method or composition for use of any one of claims 19-22, wherein the administration or delivery occurs at an interval of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days. ### Claim 24 (depends on 19, 20, 21, 22) The method or composition for use of any one of claims 19-22, wherein the administration or delivery occurs at an interval of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 weeks. ### Claim 25 (depends on 19, 20, 21, 22) The method or composition for use of any one of claims 19-22, wherein the administration or delivery occurs at an interval of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 months. ### Claim 26 The method or composition of any one of the preceding claims, wherein the guide sequence is selected from SEQ ID NOs: 5-129. ### Claim 27 The method or composition of any one of the preceding claims, wherein the guide RNA is at least partially complementary to a target sequence present in the human SERPINA1 gene. ### Claim 28 (depends on 27) The method or composition of claim 27, wherein the target sequence is in exon 2, 3, 4, or 5 of the human SERPINA1 gene. ### Claim 29 (depends on 27) The method or composition of claim 27, wherein the target sequence is in exon 2 of the human SERPINA1 gene. ### Claim 30 (depends on 27) The method or composition of claim 27, wherein the target sequence is in exon 3 of the human SERPINA1 gene. ### Claim 31 (depends on 27) The method or composition of claim 27, wherein the target sequence is in exon 4 of the human SERPINA1 gene. ### Claim 32 (depends on 27) The method or composition of claim 27, wherein the target sequence is in exon 5 of the human SERPINA1 gene. ### Claim 33 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32) The method or composition of any one of claims 1-32, wherein the guide sequence is complementary to a target sequence in the positive strand of SERPINA1. ### Claim 34 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32) The method or composition of any one of claims 1-32, wherein the guide sequence is complementary to a target sequence in the negative strand of SERPINA1. ### Claim 35 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32) The method or composition of any one of claims 1-32, further comprising a second guide sequence, wherein the first guide sequence is complementary to a first target sequence in the positive strand of the SERPINA1 gene, and wherein the second guide sequence is complementary to a second target sequence in the negative strand of the SERPINA1 gene. ### Claim 36 The method or composition of any one of the preceding claims, wherein the guide RNA comprises a crRNA that comprises the guide sequence and further comprises a nucleotide sequence of SEQ ID NO: 140, wherein the nucleotides of SEQ ID NO: 140 follow the guide sequence at its 3′ end. ### Claim 37 The method or composition of any one of the preceding claims, wherein the guide RNA is a dual guide (dgRNA). ### Claim 38 (depends on 37) The method or composition of claim 37, wherein the dual guide RNA comprises a crRNA comprising a nucleotide sequence of SEQ ID NO: 140, wherein the nucleotides of SEQ ID NO: 140 follow the guide sequence at its 3′ end, and a trRNA. ### Claim 39 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36) The method or composition of any one of claims 1-36, wherein the guide RNA is a single guide (sgRNA). ### Claim 40 (depends on 39) The method or composition of claim 39, wherein the sgRNA comprises a guide sequence that has the modifications of SEQ ID NO: 130. ### Claim 41 (depends on 39) The method or composition of claim 39, wherein the sgRNA comprises SEQ ID NO: 130. ### Claim 42 (depends on 40, 41) The method or composition of claim 40 or 41, wherein each N in SEQ ID NO: 130 is any natural or non-natural nucleotide, wherein the N's form the guide sequence, and the guide sequence targets an RNA-guided DNA binding agent to the SERPINA1 gene. ### Claim 43 (depends on 39, 40, 41, 42) The method or composition of any one of claims 39-42, wherein the sgRNA comprises any one of the guide sequences of SEQ ID NOs: 5-129 and the nucleotides of SEQ ID NO: 140. ### Claim 44 (depends on 39, 40, 41, 42, 43) The method or composition of any one of claims 39-43, wherein the sgRNA comprises a guide sequence that is at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to a sequence selected from SEQ ID Nos: 5-129. ### Claim 45 (depends on 42) The method or composition of claim 42, wherein each N in SEQ ID NO: 130 is collectively replaced with a guide sequence selected from SEQ ID Nos: 5-129. ### Claim 46 The method or composition of any one of the preceding claims, wherein the guide RNA comprises at least one modification. ### Claim 47 (depends on 46) The method or composition of claim 46, wherein the at least one modification includes a 2′-O-methyl (2′-O-Me) modified nucleotide. ### Claim 48 (depends on 46, 47) The method or composition of claim 46 or 47, wherein the at least one modification includes a phosphorothioate (PS) bond between nucleotides. ### Claim 49 (depends on 46, 47, 48) The method or composition of any one of claims 46-48, wherein the at least one modification includes a 2′-fluoro (2′-F) modified nucleotide. ### Claim 50 (depends on 46, 47, 48, 49) The method or composition of any one of claims 46-49, wherein the at least one modification includes a modification at one or more of the first five nucleotides at the 5′ end. ### Claim 51 (depends on 46, 47, 48, 49, 50) The method or composition of any one of claims 46-50, wherein the at least one modification includes a modification at one or more of the last five nucleotides at the 3′ end. ### Claim 52 (depends on 46, 47, 48, 49, 50, 51) The method or composition of any one of claims 46-51, wherein the at least one modification includes PS bonds between the first four nucleotides. ### Claim 53 (depends on 46, 47, 48, 49, 50, 51, 52) The method or composition of any one of claims 46-52, wherein the at least one modification includes PS bonds between the last four nucleotides. ### Claim 54 (depends on 46, 47, 48, 49, 50, 51, 52, 53) The method or composition of any one of claims 46-53, wherein the at least one modification includes 2′-O-Me modified nucleotides at the first three nucleotides at the 5′ end. ### Claim 55 (depends on 46, 47, 48, 49, 50, 51, 52, 53, 54) The method or composition of any one of claims 46-54, wherein the at least one modification includes 2′-O-Me modified nucleotides at the last three nucleotides at the 3′ end. ### Claim 56 (depends on 46, 47, 48, 49, 50, 51, 52, 53, 54, 55) The method or composition of any one of claims 46-55, wherein the guide RNA comprises the modified nucleotides of SEQ ID NO: 130. ### Claim 57 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56) The method or composition of any one of claims 1-56, wherein the composition further comprises a pharmaceutically acceptable excipient. ### Claim 58 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57) The method or composition of any one of claims 1-57, wherein the guide RNA and optionally the RNA-guided DNA binding agent or a nucleic acid encoding an RNA-guided DNA binding agent is/are associated with a lipid nanoparticle (LNP). ### Claim 59 (depends on 58) The method or composition of claim 58, wherein the LNP comprises a CCD lipid. ### Claim 60 (depends on 59) The method or composition of claim 59, wherein the CCD lipid is Lipid A. ### Claim 61 (depends on 58, 59, 60) The method or composition of claim 58-60, wherein the LNP comprises a neutral lipid. ### Claim 62 (depends on 61) The method or composition of claim 61, wherein the neutral lipid is DSPC ### Claim 63 (depends on 58, 59, 60, 61, 62) The method or composition of any one of claims 58-62, wherein the LNP comprises a helper lipid. ### Claim 64 (depends on 63) The method or composition of claim 63, wherein the helper lipid is cholesterol. ### Claim 65 (depends on 58, 59, 60, 61, 62, 63, 64) The method or composition of any one of claims 58-64, wherein the LNP comprises a stealth lipid. ### Claim 66 (depends on 58, 59, 60, 61, 62, 63, 64, 65) The method or composition of claim 58-65, wherein the stealth lipid is PEG2k-DMG. ### Claim 67 The method or composition of any one of the preceding claims, wherein the composition further comprises an RNA-guided DNA binding agent. ### Claim 68 The method or composition of any one of the preceding claims, wherein the composition further comprises an mRNA that encodes an RNA-guided DNA binding agent. ### Claim 69 (depends on 67, 68) The method or composition of claim 67 or 68, wherein the RNA-guided DNA binding agent is a Cas cleavase. ### Claim 70 (depends on 69) The method or composition of claim 69, wherein the RNA-guided DNA binding agent is Cas9. ### Claim 71 (depends on 67, 68, 69, 70) The method or composition of any one of claims 67-70, wherein the RNA-guided DNA binding agent is modified. ### Claim 72 (depends on 67, 68, 69, 70, 71) The method or composition of any one of claims 67-71, wherein the RNA-guided DNA binding agent is a nickase. ### Claim 73 (depends on 71, 72) The method or composition of claim 71 or 72, wherein the modified RNA-guided DNA binding agent comprises a nuclear localization signal (NLS). ### Claim 74 (depends on 67, 68, 69, 70, 71, 72, 73) The method or composition of any one of claims 67-73, wherein the RNA-guided DNA binding agent is a Cas from a Type-II CRISPR/Cas system. ### Claim 75 The method or composition of any one of the preceding claims, wherein the composition is a pharmaceutical formulation and further comprises a pharmaceutically acceptable carrier. ### Claim 76 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75) The method or composition for use of any one of claim 1-4 or 7-75, wherein the composition reduces or prevents accumulation of alpha-1 antitrypsin (AAT) in the liver. ### Claim 77 (depends on 76) The method or composition for use of claim 76, wherein the AAT is misformed. ### Claim 78 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77) The method or composition for use of any one of claim 1-4 or 7-77, wherein non-homologous ending joining (NHEJ) leads to a mutation during repair of a DSB in the SERPINA1 gene. ### Claim 79 (depends on 78) The method or composition for use of claim 78, wherein NHEJ leads to a deletion or insertion of a nucleotide(s) during repair of a DSB in the SERPINA1 gene. ### Claim 80 (depends on 79) The method or composition for use of claim 79, wherein the deletion or insertion of a nucleotide(s) induces a frame shift or nonsense mutation in the SERPINA1 gene. ### Claim 81 (depends on 80) The method or composition for use of claim 80, wherein a frame shift or nonsense mutation is induced in the SERPINA1 gene of at least 50% of liver cells. ### Claim 82 (depends on 81) The method or composition for use of claim 81, wherein a frame shift or nonsense mutation is induced in the SERPINA1 gene of 50%-60%, 60%-70%, 70% or 80%, 80%-90%, 90-95%, 95%-99%, or 99%-100% of liver cells. ### Claim 83 (depends on 79, 80, 81, 82) The method or composition for use of any one of claims 79-82, wherein a deletion or insertion of a nucleotide(s) occurs in the SERPINA1 gene at least 50-fold or more than in off-target sites. ### Claim 84 (depends on 83) The method or composition for use of claim 83, wherein the deletion or insertion of a nucleotide(s) occurs in the SERPINA1 gene 50-fold to 150-fold, 150-fold to 500-fold, 500-fold to 1500-fold, 1500-fold to 5000-fold, 5000-fold to 15000-fold, 15000-fold to 30000-fold, or 30000-fold to 60000-fold more than in off-target sites. ### Claim 85 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84) The method or composition for use of any one of claim 1-4 or 7-84, wherein administering the composition reduces levels of AAT in the subject. ### Claim 86 (depends on 85) The method or composition for use of claim 85, wherein the levels of AAT are reduced by at least 40%. ### Claim 87 (depends on 86) The method or composition for use of claim 86, wherein the levels of AAT are reduced by 40%-50%, 50%-60%, 60%-70%, 70% or 80%, 80%-90%, 90-95%, 95%-99%, or 99%-100%. ### Claim 88 (depends on 86, 87) The method or composition for use of claim 86 or 87, wherein the levels of AAT are measured in serum, plasma, blood, cerebral spinal fluid, or sputum. ### Claim 89 (depends on 86, 87) The method or composition for use of claim 86 or 87, wherein the levels of AAT are measured in liver and/or serum. ### Claim 90 (depends on 85, 86, 87, 88, 89) The method or composition for use of any one of claims 85-89, wherein the levels of AAT are measured via enzyme-linked immunosorbent assay (ELISA). ### Claim 91 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90) The method or composition for use of any one of claim 1-4 or 7-90, wherein the subject has AATD. ### Claim 92 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91) The method or composition for use of any one of claim 1-4 or 7-91, wherein the subject is human. ### Claim 93 (depends on 91, 92) The method or composition for use of claim 91 or 92, wherein the subject has AATD wt. ### Claim 94 (depends on 91, 92) The method or composition for use of claim 91 or 92, wherein the subject has hereditary AATD. ### Claim 95 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92) The method or composition for use of any one of claim 1-4, 7-92, or 94, wherein the subject has a family history of AATD. ### Claim 96 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95) The method or composition for use of any one of claim 1-4 or 7-95, wherein the subject has only or predominantly liver symptoms of AATD. ### Claim 97 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96) The method or composition for use of any one of claim 1-4 or 7-96, wherein the subject is heterozygous for the Z allele at the SERPINA1 locus. ### Claim 98 (depends on 97) The method of claim 97, wherein the subject has one Z allele and one S allele at the SERPINA1 locus. ### Claim 99 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98) The method or composition for use of any one of claim 1-4 or 7-98, wherein the subject does not have a E342K mutation in the amino acid sequence of AAT, but has reduced levels of wildtype AAT. ### Claim 100 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99) The method or composition for use of any one of claim 1-4 or 7-99, wherein the subject has an improvement, stabilization, or slowing of edema, ascites, or jaundice, or a delay in need for liver transplantation. ### Claim 101 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99) The method or composition for use of any one of claim 1-4 or 7-99, wherein the subject has an improvement, stabilization, or slowing of change as measured by imaging methods or liver enzyme levels as a result of administration. ### Claim 102 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101) The method or composition for use of any one of claim 1-4 or 7-101, wherein the composition or pharmaceutical formulation is administered via a viral vector. ### Claim 103 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102) The method or composition for use of any one of claim 1-4 or 7-102, wherein the composition or pharmaceutical formulation is administered via lipid nanoparticles. ### Claim 104 (depends on 1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103) The method or composition for use of any one of claim 1-4 or 7-103, wherein the subject is tested for specific mutations in the SERPINA1 gene before administering the composition or formulation. ### Claim 105 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 5. ### Claim 106 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 6. ### Claim 107 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 7. ### Claim 108 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 8. ### Claim 109 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 9. ### Claim 110 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 10. ### Claim 111 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 11. ### Claim 112 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 12. ### Claim 113 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 13. ### Claim 114 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 14. ### Claim 115 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 15. ### Claim 116 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 16. ### Claim 117 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 17. ### Claim 118 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 18. ### Claim 119 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 19. ### Claim 120 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 20. ### Claim 121 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 21. ### Claim 122 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 22. ### Claim 123 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 23. ### Claim 124 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 24. ### Claim 125 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 25. ### Claim 126 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 26. ### Claim 127 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 27. ### Claim 128 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 28. ### Claim 129 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 29. ### Claim 130 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 30. ### Claim 131 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 31. ### Claim 132 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 32. ### Claim 133 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 33. ### Claim 134 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 34. ### Claim 135 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 35. ### Claim 136 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 36. ### Claim 137 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 37. ### Claim 138 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 38. ### Claim 139 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 39. ### Claim 140 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 40. ### Claim 141 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 41. ### Claim 142 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 42. ### Claim 143 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 43. ### Claim 144 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 44. ### Claim 145 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 45. ### Claim 146 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 46. ### Claim 147 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 47. ### Claim 148 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 48. ### Claim 149 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 49. ### Claim 150 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 50. ### Claim 151 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 51. ### Claim 152 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 52. ### Claim 153 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 53. ### Claim 154 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 54. ### Claim 155 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 55. ### Claim 156 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 56. ### Claim 157 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 57. ### Claim 158 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 58. ### Claim 159 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 59. ### Claim 160 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 60. ### Claim 161 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 61. ### Claim 162 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 62. ### Claim 163 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 63. ### Claim 164 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 64. ### Claim 165 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 65. ### Claim 166 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 66. ### Claim 167 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 67. ### Claim 168 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 68. ### Claim 169 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 69. ### Claim 170 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 70. ### Claim 171 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 71. ### Claim 172 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 72. ### Claim 173 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 73. ### Claim 174 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 74. ### Claim 175 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 75. ### Claim 176 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 76. ### Claim 177 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 77. ### Claim 178 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 78. ### Claim 179 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 79. ### Claim 180 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 80. ### Claim 181 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 81. ### Claim 182 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 82. ### Claim 183 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 83. ### Claim 184 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 84. ### Claim 185 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 85. ### Claim 186 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 86. ### Claim 187 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 87. ### Claim 188 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 88. ### Claim 189 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 89. ### Claim 190 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 90. ### Claim 191 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 91. ### Claim 192 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 92. ### Claim 193 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 93. ### Claim 194 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 94. ### Claim 195 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 95. ### Claim 196 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 96. ### Claim 197 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 97. ### Claim 198 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 98. ### Claim 199 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 99. ### Claim 200 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 100. ### Claim 201 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 101. ### Claim 202 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 102. ### Claim 203 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 103. ### Claim 204 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 104. ### Claim 205 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 105. ### Claim 206 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 106. ### Claim 207 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 107. ### Claim 208 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 108. ### Claim 209 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 109. ### Claim 210 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 110. ### Claim 211 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 111. ### Claim 212 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 112. ### Claim 213 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 113. ### Claim 214 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 114. ### Claim 215 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 115. ### Claim 216 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 116. ### Claim 217 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 117. ### Claim 218 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 118. ### Claim 219 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 119. ### Claim 220 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 120. ### Claim 221 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 121. ### Claim 222 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 122. ### Claim 223 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 123. ### Claim 224 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 124. ### Claim 225 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 125. ### Claim 226 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 126. ### Claim 227 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 127. ### Claim 228 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 128. ### Claim 229 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 5-129 is SEQ ID NO: 129. ### Claim 230 (depends on 1, 104) The method or composition of any one of claims 1-104, further comprising the sequence of SEQ ID NO: 140 or 141. ### Claim 231 (depends on 230) The method or composition of claim 230 comprising the modification pattern of SEQ ID NO: 130. ### Claim 232 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence is selected from SEQ ID NO: 131-139. ### Claim 233 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 131. ### Claim 234 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 132. ### Claim 235 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 133. ### Claim 236 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 134. ### Claim 237 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 135. ### Claim 238 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 136. ### Claim 239 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 137. ### Claim 240 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 138. ### Claim 241 (depends on 1, 104) The method or composition of any one of claims 1-104, wherein the sequence selected from SEQ ID NOs: 131-139 is SEQ ID NO: 139. ### Claim 242 (depends on 232, 233, 234, 235, 236, 237, 238, 239, 240, 241) The method or composition of any one of claims 232-241, wherein the sequence selected from SEQ ID NOs: 131-139 comprises the modifications shown for the respective sequence in Table 2. ### Claim 243 (depends on 5, 242) Use of a composition or formulation of any of claims 5-242 for the preparation of a medicament for treating a human subject having AATD.