ENZYMES WITH RUVC DOMAINS
The present disclosure provides for endonuclease enzymes having distinguishing domain features, as well as methods of using such enzymes or variants thereof.
1 - 22 . (canceled)
23 . An engineered nuclease composition comprising:
(a) an endonuclease comprising a RuvC_III domain and an HNH domain, wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2490, 2491, 3129, 2917-2924 and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4304, 4305, 4943, 4731-4738; and
(b) an engineered guide ribonucleic acid structure configured to form a complex with said endonuclease comprising:
(i) a guide ribonucleic acid sequence configured to hybridize to a target deoxyribonucleic acid sequence; and
(ii) a tracr ribonucleic acid sequence configured to bind to said endonuclease.
24 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2490; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4304.
25 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2491; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4305.
26 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 3129; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4943.
27 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2917; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4731.
28 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2918; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4732.
29 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2919; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4733.
30 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2920; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4734.
31 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2921; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4735.
32 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2922; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4736.
33 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2923; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4737.
34 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 85% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2924; and wherein said HNH domain comprises a sequence having at least 75% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4738.
35 . The engineered nuclease composition of claim 23 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2917-2921, 2923, and 2924; and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4731-4735, 4737, and 4738.
36 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2917 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4731.
37 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2918 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4732.
38 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2919 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4733.
39 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2920 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4734.
40 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2921 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4735.
41 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2923 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4737.
42 . The engineered nuclease composition of claim 35 , wherein said RuvC_III domain comprises a sequence having at least 90% sequence identity to at least 21 consecutive amino acids of any one of SEQ ID NOs: 2924 and wherein said HNH domain comprises a sequence having at least 80% sequence identity to at least 20 consecutive amino acids of any one of SEQ ID NOs: 4738.