Engineered class 2 type v crispr systems
AssigneeScribe Therapeutics Inc.
InventorsGayathri VIJAYAKUMAR, Sean HIGGINS, Isabel COLIN, Sarah DENNY, Brett T. STAAHL, Benjamin OAKES, Angus SIDORE, Suraj MAKHIJA
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Abstract
Provided herein are engineered Class 2, Type V nucleases and guide RNAs useful for the editing of target nucleic acids. Also provided are methods of making and using such variants to modify nucleic acids.
Claims29
A gene editing pair, comprising a Class 2, Type V CRISPR protein and a guide RNA (gRNA) scaffold wherein:
- a. the gRNA scaffold comprises a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 85%, at least about 85%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity to SEQ ID NO:
- 2239 and comprises a modification at position C9, U11, U24, A29, U24, U54, G64, A88 or A95 relative to SEQ ID NO:
- 2239;
- and
- b. a gRNA comprising the gRNA scaffold and the Class 2, Type V CRISPR protein are capable of forming a ribonuclear protein complex (RNP).
A Class 2, Type V CRISPR protein comprising
- a. a NTSB domain comprising a sequence of SEQ ID NO:
- 2335, or a sequence having at least 80% at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity thereto;
- b. a helical I-II domain comprising a sequence of SEQ ID NO:
- 2336, or a sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity thereto;
- c. a helical II domain comprising a sequence of SEQ ID NO:
- 2351, or a sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity thereto;
- and
- d. a RuvC-I domain comprising a sequence of SEQ ID NO:
- 2352, or a sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity thereto.
A gRNA scaffold comprising a sequence having at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to any one of the sequences selected from the group consisting of SEQ ID NOS:
- 2292, 2291, 2307, 2281-2290, 2293-2306, 2308-2332 and 2353-2398.
Dependencies are inferred from the claim text.