	US 12,344,870 B2
	Compositions comprising a CRISPR nuclease and uses thereof
Shaorong Chong, Arlington, MA (US); Quinton Norman Wessells, Cambridge, MA (US); Roy Ziblat, Newton, MA (US); Noah Michael Jakimo, Cambridge, MA (US); Anthony James Garrity, Hingham, MA (US); and Lauren E. Alfonse, Boston, MA (US)
Assigned to Arbor Biotechnologies, Inc., Cambridge, MA (US)
Filed by Arbor Biotechnologies, Inc., Cambridge, MA (US)
Filed on Sep. 8, 2022, as Appl. No. 17/930,595.
Claims priority of provisional application 63/296,741, filed on Jan. 5, 2022.
Claims priority of provisional application 63/241,821, filed on Sep. 8, 2021.
Prior Publication US 2023/0235304 A1, Jul. 27, 2023
Int. Cl. C12N 9/22 (2006.01)

CPC C12N 9/22 (2013.01) [C12Y 301/25 (2013.01); C12N 2310/20 (2017.05)]

20 Claims

1. A variant polypeptide that comprises a substitution that corresponds to a substitution in the polypeptide of SEQ ID NO: 3 selected from D509R, S511R, I521R, D535G, Q514G, L516R, L516G, E198R, S527R, D509K, D509G, L580G, V359R, D535K, Y381R, R354G, M380K, M380R, V383R, L580R, E367R, 1521K, E367G, E507R, I521G, W350R, D535R, R528K, S527K, L385G, W350G, L516K, Y381G, H532R, D129R, P615R, G578R, M380G, V595G, R531G, D129G, R531K, D158G, N476G, G578K, A512R, P618R, V595R, V595K, V383G, A597G, Y381K, P618G, E198K, T288R, E507K, L385R, C538G, P615G, D386R, S511K, C371G, K136G, N220R, S78K, K141G, K240R, D277R, T165R, and K374R, and wherein the variant polypeptide comprises:

(a) at least 98% sequence identity to the polypeptide of SEQ ID NO: 3; or

(b) an amino acid sequence that differs from SEQ ID NO: 3 solely by the modification of 1-13 amino acids within SEQ ID NO: 3; and

wherein the variant polypeptide has nuclease activity.