CRISPR hybrid DNA/RNA polynucleotides and methods of use
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.
1. A single polynucleotide comprising:
a targeting region that comprises deoxyribonucleic acid (DNA) or ribonucleic acid (RNA); and
an activating region adjacent to said targeting region that comprises DNA; wherein said activating region comprises a stem loop structure and is capable of binding with a Cpf1.
2. The polynucleotide of claim 1 , wherein the targeting region comprises a mixture of DNA and RNA.
3. The polynucleotide of claim 1 , wherein the targeting region comprises a mixture of DNA and RNA and the activating region comprises a mixture of DNA and RNA.
4. A Class 2 CRISPR system consisting essentially of:
(i) a single polynucleotide comprising (a) a targeting region that comprises deoxyribonucleic acid (DNA) or ribonucleic acid (RNA); and (b) an activating region adjacent to said targeting region that comprises DNA; wherein said activating region comprises a stem loop structure and is capable of binding with a Cpf1; and
(ii) a Cpf1.
5. The Class 2 CRISPR system of claim 4 , wherein said targeting region comprises a mixture of DNA and RNA.
6. The Class 2 CRISPR system of claim 4 , wherein said activating region comprises a mixture of DNA and RNA.
7. The Class 2 CRISPR system of claim 4 , wherein said targeting region comprises a mixture of DNA and RNA and said activating region comprises a mixture of DNA and RNA.
8. The Class 2 CRISPR system of claim 4 , further comprising a donor polynucleotide.