Nucleobase editors and methods of use thereof
AssigneeOhio State Innovation Foundation
InventorsRenzhi Han, Li Xu, Yandi Gao
Life of this patent
Estimating the term…
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.
Claims15
A fusion protein comprising a Cas9 nickase and a monomeric adenine deaminase,
- wherein the Cas9 nickase comprises amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, and T1337R, and at least one additional mutation selected from R1335V, R1335Q or R1335E when compared to SEQ ID NO:
- 11, and
- wherein the monomeric adenine deaminase comprises amino acid substitutions A56G and V82G when compared to SEQ ID NO:
- 47.
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 monomeric adenine deaminase fused to the N-terminus of the N-terminal portion of the Cas9 nickase,
- wherein the Cas9 nickase comprises amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, and T1337R, and at least one additional mutation selected from R1335V, R1335Q or R1335E when compared to SEQ ID NO:
- 11, and
- wherein the monomeric adenine deaminase comprises amino acid substitutions A56G and V82G when compared to SEQ ID NO:
- 47.
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 monomeric adenine deaminase fused to the N-terminus of the N-terminal portion of the Cas9 nickase,
- wherein the Cas9 nickase comprises amino acid substitutions L1111R, D1135V, G1218R, E1219F, A1322R, and T1337R, and at least one additional mutation selected from R1335V, R1335Q or R1335E when compared to SEQ ID NO:
- 11, and
- wherein the monomeric adenine deaminase comprises amino acid substitutions A56G and V82G when compared to SEQ ID NO:
- 47.
Dependencies are inferred from the claim text.