	US 12,435,331 B2
	Cytosine to guanine base editor
David R. Liu, Cambridge, MA (US); and Luke W. Koblan, Cambridge, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
Filed on Nov. 28, 2022, as Appl. No. 18/059,308.
Application 18/059,308 is a division of application No. 16/492,553, granted, now 11,542,496, previously published as PCT/US2018/021878, filed on Mar. 9, 2018.
Claims priority of provisional application 62/470,175, filed on Mar. 10, 2017.
Prior Publication US 2024/0035017 A1, Feb. 1, 2024
Int. Cl. C12N 15/10 (2006.01); C07K 14/47 (2006.01); C12N 9/22 (2006.01); C12N 9/24 (2006.01); C12N 9/78 (2006.01)

CPC C12N 15/102 (2013.01) [C07K 14/47 (2013.01); C12N 9/22 (2013.01); C12N 9/2497 (2013.01); C12N 9/78 (2013.01); C07K 2319/80 (2013.01); C12Y 302/02027 (2013.01); C12Y 305/04005 (2013.01)]

29 Claims

1. A polynucleotide encoding a fusion protein comprising (i) a nucleic acid programmable DNA binding protein (napDNAbp) domain, wherein the napDNAbp domain when in association with a guide RNA (gRNA) specifically binds a target nucleic acid molecule; (ii) a cytidine deaminase domain, wherein the cytidine deaminase domain deaminates a cytosine base in the target nucleic acid molecule; and (iii) a uracil binding protein (UBP), wherein the UBP is a uracil DNA glycosylase (UDG) or a uracil base excision enzyme; wherein the fusion protein comprises the structure:

NH₂-[cytidine deaminase domain]-[napDNAbp domain]-[UBP]-COOH,

NH₂-[cytidine deaminase domain]-[UBP]-[napDNAbp domain]-COOH;

NH₂-[UBP]-[cytidine deaminase domain]-[napDNAbp domain]-COOH;

NH₂-[UBP]-[napDNAbp domain]-[cytidine deaminase domain]-COOH;

NH₂-[napDNAbp]-[UBP]-[cytidine deaminase]-COOH; or

NH₂-[napDNAbp]-[cytidine deaminase]-[UBP]-COOH;

wherein each instance of “]-[” comprises an optional linker.