US 12,264,314 B1
CasZ compositions and methods of use
Jennifer A. Doudna, Berkeley, CA (US); David Burstein, Berkeley, CA (US); Janice S. Chen, Berkeley, CA (US); Lucas B. Harrington, Berkeley, CA (US); David Paez-Espino, Walnut Creek, CA (US); and Jillian F. Banfield, Berkeley, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Filed by The Regents of the University of California, Oakland, CA (US)
Filed on Dec. 21, 2023, as Appl. No. 18/392,905.
Application 18/392,905 is a continuation of application No. 17/935,521, filed on Sep. 26, 2022, abandoned.
Application 17/935,521 is a continuation of application No. 16/896,711, filed on Jun. 9, 2020, granted, now 11,453,866, issued on Sep. 27, 2022.
Application 16/896,711 is a continuation of application No. 16/694,720, filed on Nov. 25, 2019, abandoned.
Application 16/694,720 is a continuation of application No. PCT/US2018/058545, filed on Oct. 31, 2018.
Claims priority of provisional application 62/580,395, filed on Nov. 1, 2017.
Int. Cl. C12N 15/10 (2006.01); C12N 9/10 (2006.01); C12N 15/86 (2006.01)
CPC C12N 15/102 (2013.01) [C12N 9/1007 (2013.01); C12N 15/86 (2013.01); C12N 2750/14143 (2013.01)] 30 Claims
 
1. A method of modifying a target locus of interest comprising delivering to said locus:
a) an engineered polypeptide or a nucleic acid encoding the engineered polypeptide, wherein the engineered polypeptide is a variant of the polypeptide according to SEQ ID NO: 3, wherein the variant has reduced nucleic acid cleavage activity relative to the polypeptide according to SEQ ID NO: 3, wherein the variant comprises an amino acid sequence at least 80% identical to SEQ ID NO: 3, and wherein the variant comprises at least one amino acid substitution at a position selected from a group consisting of D326, E422, R490, and D510 of SEQ ID NO: 3; and
b) a guide nucleic acid or DNA molecule encoding the guide nucleic acid, the guide nucleic acid comprising:
i) a first sequence that is bound by the polypeptide, wherein the first sequence comprises a nucleotide sequence that is at least 90% identical to residues 28-37 of SEQ ID NO: 53, and
ii) a second sequence that hybridizes to a target sequence of a target nucleic acid, wherein the first sequence is located 5′ of the second sequence.