Nucleobase editors and methods of use thereof
AssigneeOhio State Innovation Foundation
InventorsRenzhi HAN, Li XU, Yandi GAO
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Abstract
The present disclosure relates to nucleobase editors and methods of use thereof. Disclosed herein are fusion proteins, systems, and compositions for editing disease-associated mutations and methods of use thereof. In some aspects, disclosed herein is a fusion protein comprising a Cas9 nickase and a nucleotide deaminase, wherein the Cas9 nickase comprises a first amino acid substitution at a position selected from the group consisting of 262, 324, 409, 480, 543, 694, and 1219 when compared to SEQ ID NO: 11, and wherein the Cas9 nickase comprises a second amino acid substitution at a position selected from the group consisting of 1111, 1135, 1218, 1219, 1322, 1335, and 1337 when compared to SEQ ID NO: 11.
Claims31
A fusion protein comprising a Cas9 nickase and a nucleotide deaminase,
- wherein the Cas9 nickase comprises a first amino acid substitution at a position selected from the group consisting of 262, 324, 409, 480, 543, 694, and 1219 when compared to SEQ ID NO:
- 11, and
- wherein the Cas9 nickase comprises a second amino acid substitution at a position selected from the group consisting of 1111, 1135, 1218, 1219, 1322, 1335, and 1337 when compared to SEQ ID NO:
- 11.
A system for base editing comprising:
- a first nucleotide sequence encoding a N-terminal portion of a Cas9 nickase fused at its C-terminus to an intein-N;
- and
- a second nucleotide sequence encoding an intein-C fused to the N-terminus of a C-terminal portion of the Cas9 nickase;
- wherein the first nucleotide sequence further comprises a nucleotide sequence encoding a nucleotide deaminase fused to the N-terminus of the N-terminal portion of the Cas9 nickase,
- wherein the Cas9 nickase comprises a first amino acid substitution at a position selected from the group consisting of 262, 324, 409, 480, 543, 694, and 1219 when compared to SEQ ID NO:
- 11, and
- wherein the Cas9 nickase comprises a second amino acid substitution at a position selected from the group consisting of 1111, 1135, 1218, 1219, 1322, 1335, and 1337 when compared to SEQ ID NO:
- 11.
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A composition comprising:
- a first recombinant adeno-associated virus (AAV) particle comprising a first nucleotide sequence encoding a N-terminal portion of a Cas9 nickase fused at its C-terminus to an intein-N;
- and
- a second recombinant AAV particle comprising a second nucleotide sequence encoding an intein-C fused to the N-terminus of a C-terminal portion of the Cas9 nickase;
- wherein first nucleotide sequence further comprises a nucleotide sequence encoding a nucleotide deaminase fused to the N-terminus of the N-terminal portion of the Cas9 nickase,
- wherein the Cas9 nickase comprises a first amino acid substitution at a position selected from the group consisting of 262, 324, 409, 480, 543, 694, and 1219 when compared to SEQ ID NO:
- 11, and
- wherein the Cas9 nickase comprises a second amino acid substitution at a position selected from the group consisting of 1111, 1135, 1218, 1219, 1322, 1335, and 1337 when compared to SEQ ID NO:
- 11.
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Dependencies are inferred from the claim text.