US 12,460,189 B2
RNA-guided nucleases and DNA binding proteins
Neena Kenton Pyzocha, New Boston, MA (US); Adam Patrick Joyce, Stow, MA (US); and Karl Kremling, Cambridge, MA (US)
Assigned to INARI AGRICULTURE TECHNOLOGY, INC., Cambridge, MA (US)
Appl. No. 17/309,075
Filed by INARI AGRICULTURE TECHNOLOGY, INC., Cambridge, MA (US)
PCT Filed Nov. 8, 2019, PCT No. PCT/US2019/060442
§ 371(c)(1), (2) Date Apr. 21, 2021,
PCT Pub. No. WO2020/097445, PCT Pub. Date May 14, 2020.
Claims priority of provisional application 62/832,130, filed on Apr. 10, 2019.
Claims priority of provisional application 62/758,307, filed on Nov. 9, 2018.
Prior Publication US 2021/0388333 A1, Dec. 16, 2021
Int. Cl. C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/86 (2006.01)
CPC C12N 9/22 (2013.01) [C12N 15/102 (2013.01); C12N 15/86 (2013.01); C07K 2319/09 (2013.01); C07K 2319/71 (2013.01); C12N 2310/20 (2017.05)] 16 Claims
OG exemplary drawing
 
1. A composition comprising:
a) a CasP6 polypeptide, or a nucleic acid molecule encoding the CasP6 polypeptide, wherein the CasP6 polypeptide has 90% or more identity to the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3, wherein the CasP6 polypeptide comprises a RuvC-I domain, a RuvC-II domain, and a RuvC-III domain, and
wherein the RuvC-I domain comprises the amino acid sequence of SEQ ID NO: 10, 13, or 16 or one of the sequences comprising a single conservative amino acid substitution, the RuvC-II domain comprises the amino acid sequence of SEQ ID NO: 11, 14, or 17 or one of the sequences comprising a single conservative amino acid substitution, and the RuvC-III domain comprises the amino acid sequence of SEQ ID NO: 12, 15, or 18 or one of the sequences comprising a single conservative amino acid substitution, and
wherein the CasP6 polypeptide comprises catalytic amino acid residues of D434 and E617 when numbered according to SEQ ID NO: 1, D484 and E667 when numbered according to SEQ ID NO: 2, or D503 and E750 when numbered according to SEQ ID NO: 3; and
b) a recombinant CasP6 guide RNA, or one or more recombinant DNA molecules encoding the CasP6 guide RNA, wherein:
(i) the recombinant CasP6 guide RNA is capable of binding to a CasP6 protein, forming a ribonucleoprotein complex (RNP), and targeting the complex to a specific location within a target single stranded RNA (ssRNA); and
(ii) the CasP6 polypeptide and the recombinant CasP6 guide RNA, or one or more recombinant DNA molecules encoding the recombinant CasP6 guide RNA, do not naturally occur together.