Methods and compositions for directed genome editing
AssigneeRewrite Therapeutics, Inc.
InventorSchaked Omer HALPERIN
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Abstract
Provided herein are compositions and methods for increasing editing efficiency of a target nucleic acid. A composition may comprise a guide nucleic acid, a Cas9 nickase, or a reverse transcriptase. The reverse transcriptase may be fused to the Cas9 nickase. The reverse transcriptase may heterodimerize with the Cas9 nickase. The reverse transcriptase may bind to a guide nucleic acid. The reverse transcriptase may be engineered to increase processivity. The guide nucleic acid may be engineered to facilitate synthesis or editing of a sequence. The guide nucleic acid, Cas9 nickase, and reverse transcriptase may be engineered to fit within AAV vectors. The guide nucleic acid may comprise a region that binds to another region on the guide nucleic acid to improve gene editing.
Claims21
1-131. (canceled)
A method, comprising:
- contacting a target nucleic acid with one or more nucleic acids collectively comprising:
- a spacer reverse complementary to a first region of the target nucleic acid;
- a scaffold configured to bind to a nuclease;
- a reverse transcriptase template encoding a sequence to be inserted into the target nucleic acid;
- a primer binding site reverse complementary to a second region of the target nucleic acid;
- and
- at least one of (i) or (ii):
- (i) a guide nucleic acid positioning system (GPS) region and a GPS binding site that hybridizes to the GPS region, wherein the GPS region and the GPS binding site are at least 10 nucleotides in length and are at least 60% reverse complementary to each other, or
- (ii) a modification in the reverse transcriptase template that disrupts a track of at least 4 consecutive nucleotides of the same base in the target nucleic acid.
Dependencies are inferred from the claim text.