Compositions and Methods for Treating Alpha-1 Antitrypsin Deficiency

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.

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.

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.

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.

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.

The method or composition of any one of the preceding claims, wherein the guide sequence is selected from SEQ ID NOs:

• 5-129.

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.

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.

The method or composition of any one of the preceding claims, wherein the guide RNA is a dual guide (dgRNA).

The method or composition of any one of the preceding claims, wherein the guide RNA comprises at least one modification.

The method or composition of any one of the preceding claims, wherein the composition further comprises an RNA-guided DNA binding agent.

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.

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.