Granted Patent B2

US 12,390,538 · App. 18/762,259 · Granted Aug 19, 2025

Inventors: Aron Brandon Jaffe (Brookline, MA); Noorussahar Abubucker (Watertown, MA); Yesseinia Anglero-Rodriguez (Everett, MA); Vic Myer (Arlington, MA); Angelo Leone Lombardo (Rome, IT); Martino Alfredo Cappelluti (Milan, IT)

Assignee: nChroma Bio, Inc.

A61K48/005A61P31/20C12N9/1007C12N9/22C12N15/11C12Y201/01037C07K2319/00C07K2319/80C12N2310/20

Abstract

Claims (25)

1. A method, comprising administering an epigenetic editing system to a subject, wherein the subject comprises detectable levels of HBV DNA, HBsAg, and/or HBeAg in plasma of the subject, wherein the epigenetic editing system comprises

a) a fusion protein comprising

i) a dCas9 protein domain,

ii) a DNMT3A domain and

iii) a human KRAB domain

or one or more nucleic acid molecules encoding the fusion protein,

and b) a guide RNA (gRNA) comprising a nucleic acid base sequence selected from the group consisting of SEQ ID NOs: 1093-1110, 1130-1164, 1166-1173, 1175-1199, 1201-1235, and 1249, wherein the nucleic acid base sequence comprises a region complementary to a strand of a target region wherein the target region comprises a sequence selected from the group consisting of SEQ ID NOs: 333-350, 370-404, 406-413, 415-439, and 441-475.

2. The method of claim 1 , wherein the fusion protein further comprises a DNMT3L domain.

3. The method of claim 1 , wherein the dCas9 protein domain is from Streptococcus pyogenes or Staphylococcus aureus.

4. The method of claim 3 , wherein the dCas9 protein domain comprises a sequence selected from the group consisting of SEQ ID NOs: 12, 13, 27, 28 and 29.

5. The method of claim 3 , wherein the dCas9 protein domain is from Streptococcus pyogenes.

6. The method of claim 4 , wherein the dCas9 protein domain comprises the sequence of SEQ ID NO: 12.

7. The method of claim 1 , wherein the DNMT3A domain comprises a sequence selected from the group consisting of SEQ ID NOs: 1028 and 1029.

8. The method of claim 1 , wherein the DNMT3A domain comprises a sequence of SEQ ID NO: 1029.

9. The method of claim 1 , wherein the human KRAB domain comprises a sequence selected from the group consisting of SEQ ID NOs: 495, 551, 536, 537, 707, and 717.

10. The method of claim 1 , wherein the human KRAB domain is a ZIM3 KRAB domain.

11. The method of claim 10 , wherein the ZIM3 KRAB domain comprises the sequence of SEQ ID NO: 495.

12. The method of claim 1 , wherein the target region comprises the sequence of SEQ ID NO: 391.

13. The method of claim 1 , wherein the target region comprises the sequence of SEQ ID NO: 392.

14. The method of claim 1 , wherein the gRNA comprises the nucleic acid base sequence of SEQ ID NO: 1151 or 1249.

15. The method of claim 2 , wherein the DNMT3L domain is a human DNMT3L domain.

16. The method of claim 2 , wherein the DNMT3L domain comprises a sequence selected from the group consisting of SEQ ID NOs: 1032-1035.

17. The method of claim 16 , wherein the DNMT3L domain comprises the sequence of SEQ ID NO: 1033.

18. The method of claim 1 , wherein the dCas9 protein domain comprises a sequence of SEQ ID NO: 12, the DNMT3A domain comprises a sequence of SEQ ID NO: 1029, and the human KRAB domain comprises a sequence of SEQ ID NO: 495.

19. The method of claim 1 , wherein the dCas9 protein domain comprises a sequence with at least 99% sequence identity to the sequence of SEQ ID NO: 12, the DNMT3A domain comprises a sequence with at least 99% sequence identity to the sequence of SEQ ID NO: 1029, and the human KRAB domain comprises a sequence with at least 99% sequence identity to the sequence of SEQ ID NO: 495.

Assignments (9)

Continuity (5)

References Cited (400)