	US 12,234,453 B2
	Regulation of transcription through CTCF loop anchors
Jurian Schuijers, Somerville, MA (US); Abraham S. Weintraub, Cambridge, MA (US); John C. Manteiga, Boston, MA (US); and Richard A. Young, Boston, MA (US)
Assigned to Whitehead Institute for Biomedical Research, Cambridge, MA (US)
Filed by Whitehead Institute for Biomedical Research, Cambridge, MA (US)
Filed on Jun. 5, 2023, as Appl. No. 18/329,541.
Application 18/329,541 is a division of application No. 16/469,131, abandoned, previously published as PCT/US2017/065918, filed on Dec. 12, 2017.
Claims priority of provisional application 62/433,234, filed on Dec. 12, 2016.
Prior Publication US 2024/0141335 A1, May 2, 2024
Int. Cl. C12N 15/11 (2006.01); A61K 38/00 (2006.01); C12N 9/22 (2006.01); C12N 15/90 (2006.01); C12Q 1/6886 (2018.01)

CPC C12N 15/11 (2013.01) [C12N 9/22 (2013.01); C12N 15/907 (2013.01); C12Q 1/6886 (2013.01); A61K 38/00 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01); C12Q 2600/106 (2013.01); C12Q 2600/154 (2013.01)]

20 Claims

1. A method for methylating a promoter region CTCF binding site of a gene in a cell comprising introducing into the cell:

(a) a fusion protein comprising a catalytically inactive Cas9 operably linked to at least one DNA methyltransferase, or a nucleic acid encoding the fusion protein; and

(b) a nucleic acid comprising one or more guide sequences homologous to a sequence, wherein the sequence is (i) in the promoter region CTCF binding site or (ii) within a region of 100, 200, 300, 400, 500, 600, 700, 800, 900 or 1000 nucleotides upstream or downstream of the promoter region CTCF binding site, thereby methylating the promoter region CTCF binding site.