# US 2025/0154480 A1 · Treatment for nucleotide repeat expansion disease > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0154480 A1 - Document ID: US20250154480A1 - Status: Published application - Kind: A1 - Application number: 18/838,002 - Filed: 2023-02-22 - Published: 2025-05-15 - Earliest claimed: 2022-02-23 - Assignees: The Johns Hopkins University - Applicants: The Johns Hopkins University - Inventors: Jiou Wang; Honghe Liu - Web page: https://patentgrep.com/patent/US20250154480A1 ## CPC - C12N 9/226: {Class 2 CAS enzyme complex, e.g. single CAS protein} - A61K 38/465: {acting on ester bonds (3.1), e.g. lipases, ribonucleases} - A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered} - A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia - C07K 14/47: from mammals - C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} - 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)} - C12N 15/52: Genes encoding for enzymes or proenzymes - C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR] - C12N 2320/30: Special therapeutic applications - C12N 2510/00: Genetically modified cells - 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 - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract Provided herein is technology relating to nucleotide repeat elements present in the genomes of eukaryotes and particularly, but not exclusively, to technologies for reducing the levels of disease-causing products expressed from expansions of nucleotide repeats. ## Claims ### Claim 1 A ribonucleoprotein (RNP) comprising a Cas13 protein and a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 2 (depends on 1) The RNP of claim 1, wherein said Cas13 protein is Cas13d. ### Claim 3 (depends on 1) The RNP of claim 1, wherein said spacer sequence is 24 nt long. ### Claim 4 (depends on 1) The RNP of claim 1, wherein said spacer sequence is 30 nt long. ### Claim 5 (depends on 1) The RNP of claim 1, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 6 (depends on 1) The RNP of claim 1, wherein said gRNA comprises a sequence provided by SEQ TD NO: 8. ### Claim 7 (depends on 1) The RNP of claim 1, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 8 A nucleic acid comprising a first nucleotide sequence encoding a Cas13 protein and a second nucleotide sequence encoding a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 9 (depends on 8) The nucleic acid of claim 8, wherein said Cas13 protein is Cas13d. ### Claim 10 (depends on 8) The nucleic acid of claim 8, wherein said spacer sequence is 24 nt long. ### Claim 11 (depends on 8) The nucleic acid of claim 8, wherein said spacer sequence is 30 nt long. ### Claim 12 (depends on 8) The nucleic acid of claim 8, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 13 (depends on 8) The nucleic acid of claim 8, wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 14 (depends on 8) The nucleic acid of claim 8, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 15 (depends on 8) A vector comprising the nucleic acid of claim 8. ### Claim 16 (depends on 8) A cell comprising the nucleic acid of claim 8. ### Claim 17 (depends on 16) The cell of claim 16, wherein a vector comprises the nucleic acid. ### Claim 18 (depends on 16) The cell of claim 16, wherein the cell is a human cell comprising a hexanucleotide repeat in chromosome 9. ### Claim 19 (depends on 16) The cell of claim 16, wherein the cell is a human cell comprising a hexanucleotide repeat at a C9orf72 locus. ### Claim 20 (depends on 16) The cell of claim 16, wherein the cell is a human cell comprising one or more GGGGCC repeats at a C9orf72 locus. ### Claim 21 (depends on 16) The cell of claim 16, wherein a patient having a neurological disease comprises the cell. ### Claim 22 (depends on 16) The cell of claim 16, wherein a patient having amyotrophic lateral sclerosis comprises the cell. ### Claim 23 (depends on 16) The cell of claim 16, wherein a patient having frontotemporal dementia comprises the cell. ### Claim 24 (depends on 16) The cell of claim 16, wherein a patient having Alzheimer's disease comprises the cell. ### Claim 25 (depends on 16) The cell of claim 16, wherein a patient having Huntington's disease, multiple system atrophy, depressive pseudo dementia, or bipolar disorder comprises the cell. ### Claim 26 (depends on 1) A cell comprising the RNP of claim 1. ### Claim 27 (depends on 26) The cell of claim 26, wherein the cell is a human cell comprising a hexanucleotide repeat in chromosome 9. ### Claim 28 (depends on 26) The cell of claim 26, wherein the cell is a human cell comprising a hexanucleotide repeat at a C9orf72 locus. ### Claim 29 (depends on 26) The cell of claim 26, wherein the cell is a human cell comprising one or more GGGGCC repeats at a C9orf72 locus. ### Claim 30 (depends on 26) The cell of claim 26, wherein a patient having a neurological disease comprises the cell. ### Claim 31 (depends on 26) The cell of claim 26, wherein a patient having amyotrophic lateral sclerosis comprises the cell. ### Claim 32 (depends on 26) The cell of claim 26, wherein a patient having frontotemporal dementia comprises the cell. ### Claim 33 (depends on 26) The cell of claim 26, wherein a patient having Alzheimer's disease comprises the cell. ### Claim 34 (depends on 26) The cell of claim 26, wherein a patient having Huntington's disease, multiple system atrophy, depressive pseudo dementia, or bipolar disorder comprises the cell. ### Claim 35 A system comprising a Cas13 protein and a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 36 (depends on 35) The system of claim 35, wherein said Cas13 protein is Cas13d. ### Claim 37 (depends on 35) The system of claim 35, wherein said spacer sequence is 24 nt long. ### Claim 38 (depends on 35) The system of claim 35, wherein said spacer sequence is 30 nt long. ### Claim 39 (depends on 35) The system of claim 35, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 40 (depends on 35) The system of claim 35, wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 41 (depends on 35) The system of claim 35, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 42 A system comprising a nucleic acid encoding a Cas13 protein and a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 43 (depends on 42) The system of claim 42, wherein said Cas13 protein is Cas13d. ### Claim 44 (depends on 42) The system of claim 42, wherein said spacer sequence is 24 nt long. ### Claim 45 (depends on 42) The system of claim 42, wherein said spacer sequence is 30 nt long. ### Claim 46 (depends on 42) The system of claim 42, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 47 (depends on 42) The system of claim 42 wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 48 (depends on 42) The system of claim 42, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 49 A system comprising a Cas13 protein and a nucleic acid encoding a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 50 (depends on 49) The system of claim 49, wherein said Cas13 protein is Cas13d. ### Claim 51 (depends on 49) The system of claim 49, wherein said spacer sequence is 24 nt long. ### Claim 52 (depends on 49) The system of claim 49, wherein said spacer sequence is 30 nt long. ### Claim 53 (depends on 49) The system of claim 49, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 54 (depends on 49) The system of claim 49, wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 55 (depends on 49) The system of claim 49, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 56 A system comprising a nucleic acid encoding a Cas13 protein and a nucleic acid encoding a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 57 (depends on 56) The system of claim 56, wherein said Cas13 protein is Cas13d. ### Claim 58 (depends on 56) The system of claim 56, wherein said spacer sequence is 24 nt long. ### Claim 59 (depends on 56) The system of claim 56, wherein said spacer sequence is 30 nt long. ### Claim 60 (depends on 56) The system of claim 56, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 61 (depends on 56) The system of claim 56, wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 62 (depends on 56) The system of claim 56, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 63 A method of treating a subject having a neurological disease, said method comprising administering a ribonucleoprotein (RNP) to said subject, wherein said RNP comprises a Cas13 protein and a gRNA and wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 64 (depends on 63) The method of claim 63, wherein said Cas13 protein is Cas13d. ### Claim 65 (depends on 63) The method of claim 63, wherein said spacer sequence is 24 nt long. ### Claim 66 (depends on 63) The method of claim 63, wherein said spacer sequence is 30 nt long. ### Claim 67 (depends on 63) The method of claim 63, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 68 (depends on 63) The method of claim 63, wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 69 (depends on 63) The method of claim 63, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 70 (depends on 63) The method of claim 63, wherein said neurological disease is amyotrophic lateral sclerosis. ### Claim 71 (depends on 63) The method of claim 63, wherein said neurological disease is frontotemporal dementia. ### Claim 72 (depends on 63) The method of claim 63, wherein said neurological disease is Alzheimer's disease. ### Claim 73 (depends on 63) The method of claim 63, wherein said neurological disease is Huntington's disease, multiple system atrophy, depressive pseudo dementia, or bipolar disorder. ### Claim 74 (depends on 63) The method of claim 63, wherein said subject comprises a cell comprising a hexanucleotide repeat in chromosome 9. ### Claim 75 (depends on 63) The method of claim 63, wherein said subject comprises a cell comprising a hexanucleotide repeat at a C9orf72 locus. ### Claim 76 (depends on 63) The method of claim 63, wherein said subject comprises a cell comprising one or more GGGGCC repeats at a C9orf72 locus. ### Claim 77 A method of treating a subject having a neurological disease, said method comprising administering a nucleic acid to said subject, wherein said nucleic acid comprises a first nucleotide sequence encoding a Cas13 protein and a second nucleotide sequence encoding a gRNA and wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences. ### Claim 78 (depends on 78) The method of claim 78, wherein said Cas13 protein is Cas13d. ### Claim 79 (depends on 78) The method of claim 78, wherein said spacer sequence is 24 nt long. ### Claim 80 (depends on 78) The method of claim 78, wherein said spacer sequence is 30 nt long. ### Claim 81 (depends on 78) The method of claim 78, wherein said gRNA sequence comprises a G or A at the +1 position. ### Claim 82 (depends on 78) The method of claim 78, wherein said gRNA comprises a sequence provided by SEQ ID NO: 8. ### Claim 83 (depends on 78) The method of claim 78, wherein said gRNA comprises a sequence provided by SEQ ID NO: 9. ### Claim 84 (depends on 78) The method of claim 78, wherein said neurological disease is amyotrophic lateral sclerosis. ### Claim 85 (depends on 78) The method of claim 78, wherein said neurological disease is frontotemporal dementia. ### Claim 86 (depends on 78) The method of claim 78, wherein said neurological disease is Alzheimer's disease. ### Claim 87 (depends on 78) The method of claim 78, wherein said neurological disease is Huntington's disease, multiple system atrophy, depressive pseudo dementia, or bipolar disorder. ### Claim 88 (depends on 78) The method of claim 78, wherein said subject comprises a cell comprising a hexanucleotide repeat in chromosome 9. ### Claim 89 (depends on 78) The method of claim 78, wherein said subject comprises a cell comprising a hexanucleotide repeat at a C9orf72 locus. ### Claim 90 (depends on 78) The method of claim 78, wherein said subject comprises a cell comprising one or more GGGGCC repeats at a C9orf72 locus.