US 12,252,705 B2
Small type II-D Cas proteins and methods of use thereof
Feng Zhang, Cambridge, MA (US); Soumya Kannan, Cambridge, MA (US); and Han Altae-Tran, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed on Sep. 19, 2023, as Appl. No. 18/470,189.
Application 18/470,189 is a continuation of application No. 17/793,115, previously published as PCT/US2021/013753, filed on Jan. 15, 2021.
Claims priority of provisional application 62/962,672, filed on Jan. 17, 2020.
Prior Publication US 2024/0026382 A1, Jan. 25, 2024
Int. Cl. C12N 15/90 (2006.01); C12N 9/22 (2006.01); C12N 9/78 (2006.01); C12N 15/10 (2006.01)
CPC C12N 15/90 (2013.01) [C12N 9/22 (2013.01); C12N 9/78 (2013.01); C12N 15/102 (2013.01); C12Y 207/07049 (2013.01); C12Y 305/04004 (2013.01); C12Y 305/04005 (2013.01)] 21 Claims
 
1. An engineered nucleic acid targeting system comprising:
a Cas polypeptide comprising a RuvC domain and an HNH domain, wherein the Cas polypeptide is about 950 amino acids or less in size,
an engineered guide molecule comprising a guide sequence and a scaffold and capable of forming a complex with the Cas polypeptide and directing sequence specific binding of the complex to a target polynucleotide, and
wherein the Cas polypeptide is encoded by nucleotides 9364-12198 of SEQ ID NO: 99, nucleotides 9364-12198 of SEQ ID NO: 100; nucleotides 10001-12841 of SEQ ID NO: 101, or nucleotides 1597-4407 of SEQ ID NO: 126, or has at least 90% amino acid sequence identity to the Cas polypeptide encoded by nucleotides 9364-12198 of SEQ ID NO: 99, nucleotides 9364-12198 of SEQ ID NO: 100; nucleotides 10001-12841 of SEQ ID NO: 101, or nucleotides 1597-4407 of SEQ ID NO: 126.