US 11,939,607 B2
Gene editing systems comprising an RNA guide targeting lactate dehydrogenase a (LDHA) and uses thereof
Quinton Norman Wessells, Cambridge, MA (US); Jeffrey Raymond Haswell, Needham, MA (US); Tia Marie Ditommaso, Newton, MA (US); Noah Michael Jakimo, San Francisco, CA (US); and Sejuti Sengupta, Ashland, MA (US)
Assigned to ARBOR BIOTECHNOLOGIES, INC., Cambridge, MA (US)
Filed by Arbor Biotechnologies, Inc., Cambridge, MA (US)
Filed on Nov. 4, 2022, as Appl. No. 18/052,791.
Application 18/052,791 is a continuation of application No. 17/832,114, filed on Jun. 3, 2022.
Claims priority of provisional application 63/300,743, filed on Jan. 19, 2022.
Claims priority of provisional application 63/292,912, filed on Dec. 22, 2021.
Claims priority of provisional application 63/225,214, filed on Jul. 23, 2021.
Claims priority of provisional application 63/197,067, filed on Jun. 4, 2021.
Prior Publication US 2023/0212540 A1, Jul. 6, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/22 (2006.01); A61K 31/7105 (2006.01); A61K 38/46 (2006.01); A61K 47/54 (2017.01); A61P 13/12 (2006.01); C12N 9/04 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01); C12N 15/86 (2006.01); C12N 15/90 (2006.01)
CPC C12N 9/22 (2013.01) [A61K 31/7105 (2013.01); A61K 38/465 (2013.01); A61K 47/549 (2017.08); A61P 13/12 (2018.01); C12N 9/0006 (2013.01); C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 15/86 (2013.01); C12N 15/907 (2013.01); C12Y 101/01027 (2013.01); C12N 2310/20 (2017.05); C12N 2320/30 (2013.01); C12N 2750/14143 (2013.01); C12N 2800/80 (2013.01)] 21 Claims
 
1. A gene editing system for genetic editing of a lactate dehydrogenase A (LDHA) gene, comprising
(i) a Cas12i2 polypeptide or a first nucleic acid encoding the Cas12i2 polypeptide, wherein the Cas12i2 polypeptide is a variant of SEQ ID NO:1166, and wherein, relative to SEQ ID NO: 1166, the Cas12i2 polypeptide comprises mutations at positions comprising D581, 1926, and V1030; and wherein the mutations are amino acid substitutions of D581R, I926R, and V1030G, respectively;
(ii) an RNA guide or a second nucleic acid encoding the RNA guide, wherein the RNA guide comprises a spacer sequence specific to a target sequence within an LDHA gene, the target sequence being adjacent to a protospacer adjacent motif (PAM) comprising the motif of 5′-TTN-3′, which is located 5′ to the target sequence.