	US 12,252,692 B2
	Compositions, systems, and methods for regulation of hepatitis B virus through targeted gene repression
Brian Cosgrove, Durham, NC (US); Kendra Congdon, Durham, NC (US); Jason Dean, Seattle, WA (US); Veronica Gough, Durham, NC (US); Joshua B. Black, Durham, NC (US); and Britta Jones, Durham, NC (US)
Assigned to Tune Therapeutics, Inc., Seattle, WA (US)
Filed by Tune Therapeutics, Inc., Seattle, WA (US)
Filed on Aug. 18, 2023, as Appl. No. 18/452,514.
Claims priority of provisional application 63/531,309, filed on Aug. 7, 2023.
Claims priority of provisional application 63/472,236, filed on Jun. 9, 2023.
Claims priority of provisional application 63/399,634, filed on Aug. 19, 2022.
Prior Publication US 2024/0067969 A1, Feb. 29, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/113 (2010.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/88 (2006.01)

CPC C12N 15/1131 (2013.01) [C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/88 (2013.01); C12N 2310/11 (2013.01); C12N 2310/20 (2017.05); C12N 2310/315 (2013.01); C12N 2310/321 (2013.01); C12N 2310/351 (2013.01)]

25 Claims

1. An epigenetic-modifying DNA-targeting system comprising a DNA-targeting module for repressing transcription of one or more Hepatitis B viral (HBV) genes, wherein the DNA-targeting module comprises:

(a) a gRNA for targeting to a target site in a Hepatitis B viral DNA sequence, wherein the target site comprises the sequence set forth in SEQ ID NO: 22; and

(b) a fusion protein comprising a deactivated Streptococcus pyogenes Cas9 (dSpCas9) protein and a transcriptional repressor effector domain comprising a KRAB domain and a DNMT3A/L domain, wherein the KRAB domain comprises the sequence set forth in SEQ ID NO: 590 and the DNMT3A/L domain comprises the sequence set forth in SEQ ID NO: 651.