COMPOSITIONS AND METHODS FOR THE TREATMENT OF HEREDITARY ANGIOEDEMA (HAE)

1 . A method of editing a nucleobase of a kallikrein B1 (KLKB1) polynucleotide in a cell, the method comprising contacting the KLKB1 polynucleotide with one or more guide polynucleotides and a base editor comprising a fusion protein or a protein complex comprising a nucleic acid programmable DNA binding protein (napDNAbp) domain and a deaminase domain, or a polynucleotide encoding the base editor, wherein said guide polynucleotide targets said base editor to effect an alteration of the nucleobase of the KLKB1 polynucleotide.

2 . A method of treating hereditary angioedema in a subject in need thereof, the method comprising administering to a cell of the subject a base editor comprising a fusion protein or protein complex comprising a nucleic acid programmable DNA binding protein (napDNAbp) domain and a deaminase domain, or a polynucleotide encoding the base editor, and a guide polynucleotide that targets the base editor to effect an alteration of a nucleobase of a KLKB1 polynucleotide, thereby treating hereditary angioedema in the subject.

3 . The method of claim 1 , wherein the guide polynucleotide comprises one or more of the spacers listed in Table 1A or Table 1B.

4 . The method of claim 1 , wherein the deaminase is an adenosine deaminase or a cytidine deaminase.

5 . The method of claim 1 , wherein the target base editing efficiency is between at least about 20-50%.

6 . The method of claim 1 , wherein alteration of the nucleobase is associated with a reduction in transcription of a polynucleotide sequence encoding the KLKB1 protein.

7 . The method of claim 1 , wherein alteration of the nucleobase in the KLKB1 polynucleotide results in a missense mutation and/or disrupts a splice site.

8 . The method of claim 1 , wherein the base editor is selected from the group consisting of ABE8.8, IBE3, IBE6, ABE8.8-MQKFRAER, ABE8.8-VRQR, IBE3-MQKFRAER, and IBE6-VRQR.

9 . The method of claim 1 , wherein the guide polynucleotide comprises a spacer corresponding to or comprising a guide polynucleotide selected from the group consisting of gRNA1074, gRNA1090, gRNA1104, gRNA1108, gRNA1115, gRNA1150, gRNA1151, gRNA1158, gRNA1159, gRNA1166, or gRNA1178.

10 . A modified cell comprising an alteration in a nucleobase of an KLKB1 polynucleotide, wherein the alteration is prepared by the method of claim 1 , and wherein the alteration reduces or eliminates expression and/or function of the encoded KLKB1 polypeptide as compared to a control cell without the modification.

11 . A base editor system comprising a fusion protein or a polynucleotide encoding the fusion protein, wherein the fusion protein comprises a nucleic acid programmable DNA binding protein domain (napDNAbp), a deaminase domain, and a guide polynucleotide that comprises at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of a spacer listed in Table 1A or Table 1B.

12 . The base editor system of claim 11 , wherein the guide polynucleotide comprises a spacer corresponding to or comprising a guide polynucleotide selected from the group consisting of gRNA1074, gRNA1090, gRNA1104, gRNA1108, gRNA1115, gRNA1150, gRNA1151, gRNA1158, gRNA1159, gRNA1166, or gRNA1178.

13 . A polynucleotide encoding the base editor system of claim 11 , or a component thereof.

14 . A cell produced by the method of claim 1 .

15 . A kit comprising a base editor system comprising a fusion protein or a polynucleotide encoding the fusion protein, wherein the fusion protein comprises a nucleic acid programmable DNA binding protein domain (napDNAbp), a deaminase domain, and a guide polynucleotide that comprises least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of a spacer sequence listed in Table 1A or Table 1B.

16 . The kit of claim 15 , wherein the guide polynucleotide comprises a spacer corresponding to or comprising a guide polynucleotide selected from the group consisting of gRNA1074, gRNA1090, gRNA1104, gRNA1108, gRNA1115, gRNA1150, gRNA1151, gRNA1158, gRNA1159, gRNA1166, or gRNA1178.

17 . The kit of claim 15 , wherein the base editor is selected from the group consisting of ABE8.8, IBE3, IBE6, ABE8.8-MQKFRAER, ABE8.8-VRQR, IBE3-MQKFRAER, and IBE6-VRQR.

18 . A pharmaceutical composition comprising an effective amount of a base editor system comprising a fusion protein or a polynucleotide encoding the fusion protein, wherein the fusion protein comprises a nucleic acid programmable DNA binding protein domain (napDNAbp), a deaminase domain, and a guide polynucleotide that comprises at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases of a spacer listed in Table 1A or Table 1B.

19 . A guide polynucleotide comprising a sequence listed in Table 1A or Table 1B.

20 . A method of modifying an KLKB1 polynucleotide in a cell, the method comprising contacting the KLKB1 polynucleotide with a nucleic acid programmable DNA binding protein domain (napDNAbp) having endonuclease activity on both strands of a double-stranded DNA molecule and a guide polynucleotide, wherein said guide polynucleotide targets said napDNAbp to cleave the KLKB1 polynucleotide, thereby altering the expression or activity of an encoded gene product.

21 . A method of treating hereditary angioedema in a subject in need thereof, the method comprising administering to a cell of the subject with a nucleic acid programmable DNA binding protein domain (napDNAbp) having endonuclease activity and a guide polynucleotide, wherein said guide polynucleotide targets said napDNAbp to cleave the KLKB1 polynucleotide, thereby treating hereditary angioedema in the subject.

22 . The method of claim 21 , wherein the guide polynucleotide comprises a spacer sequence selected from those listed in Table 1A or Table 1B.