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US 11,236,364 B2Granted
CRISPR hybrid DNA/RNA polynucleotides and methods of use
AssigneeCaribou Biosciences, Inc.
InventorsAndrew Paul May, Paul Daniel Donohue
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Abstract
The present disclosure provides DNA-guided CRISPR systems; polynucleotides comprising DNA, RNA and mixtures thereof for use with CRISPR systems; and methods of use involving such polynucleotides and DNA-guided CRISPR systems.
Claims21
A single Class 2 CRISPR polynucleotide comprising:
- a targeting region that comprises deoxyribonucleic acid (DNA), and an activating region that comprises a mixture of DNA and ribonucleic acid (RNA);
- wherein the activating region is adjacent to the targeting region;
- and wherein the activating region comprises a stem loop structure and is capable of binding with a Cpf1 protein.
A method of modifying a target nucleic acid molecule comprising:
- contacting the target nucleic acid molecule having a target sequence with
- (i) a single polynucleotide comprising (a) a targeting region that comprises deoxyribonucleic acid (DNA), and an activating region adjacent to the targeting region, said activating region comprising a mixture of DNA and ribonucleic acid (RNA);
- or (b) a targeting region that comprises DNA or RNA and an activating region adjacent to the targeting region, said activating region comprising DNA;
- wherein the targeting region is configured to hybridize with the target sequence, and wherein the activating region comprises a stem loop structure;
- and
- (ii) a Cpf1,
- wherein the Cpf1 binds with the activating region of the single polynucleotide, wherein the contacting of the target nucleic acid molecule with the single polynucleotide and the Cpf1 occurs (A) in vitro, (B) in a non-human cell, or (C) in an isolated cell and wherein the target nucleic acid molecule is cleaved.
4 more elements
Dependencies are inferred from the claim text.