US 12,454,694 B2
Compositions and methods for improving base editing
John Evans, Cambridge, MA (US); and Jason Michael Gehrke, Cambridge, MA (US)
Assigned to Beam Therapeutics Inc., Cambridge, MA (US)
Appl. No. 17/273,071
Filed by Beam Therapeutics Inc., Cambridge, MA (US)
PCT Filed Sep. 7, 2019, PCT No. PCT/US2019/050112
§ 371(c)(1), (2) Date Mar. 3, 2021,
PCT Pub. No. WO2020/051562, PCT Pub. Date Mar. 12, 2020.
Claims priority of provisional application 62/728,698, filed on Sep. 7, 2018.
Prior Publication US 2022/0127622 A1, Apr. 28, 2022
Int. Cl. C12N 15/62 (2006.01); C12N 9/22 (2006.01); C12N 9/78 (2006.01); C12N 15/11 (2006.01); C12N 15/86 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/62 (2013.01) [C12N 9/22 (2013.01); C12N 9/78 (2013.01); C12N 15/11 (2013.01); C12N 15/86 (2013.01); C12N 15/907 (2013.01); C12Y 305/04004 (2013.01); C12Y 305/04005 (2013.01); C07K 2319/09 (2013.01); C07K 2319/80 (2013.01); C12N 2310/20 (2017.05); C12N 2710/16143 (2013.01); C12N 2800/80 (2013.01)] 17 Claims
OG exemplary drawing
 
1. A nucleobase editor system comprising
(a) a first nucleic acid programmable DNA binding protein (napDNAbp) domain that lacks nuclease activity and has nucleic acid sequence specific binding activity,
(b) a second napDNAbp domain that has nickase activity and nucleic acid sequence specific binding activity, and
(c) an adenosine deaminase domain having base editing activity and comprising an amino acid sequence having at least 95% identity to the following amino acid sequence:
(SEQ ID NO: 89)
SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIG
 
LHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRI
 
GRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYF
 
FRMPRQVFNAQKKAQSSTD,
wherein the first napDNAbp domain is complexed with a first guide RNA, wherein the second napDNAbp domain is complexed with a second guide RNA,
wherein the first napDNAbp and the second napDNAbp are each derived from different species, and
wherein the first guide RNA and the second guide RNA are different.