gRNA STABILIZATION IN NUCLEIC ACID-GUIDED NICKASE EDITING
AssigneeInscripta, Inc.
InventorAamir Mir
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Abstract
The present disclosure provides compositions of matter, methods and instruments for nucleic acid-guided nickase/reverse transcriptase fusion editing in live cells. Editing efficiency is improved using fusion proteins (e.g., the nickase-RT fusion) that retain certain characteristics of nucleic acid-directed nucleases (e.g., the binding specificity and ability to cleave one or more DNA strands in a targeted manner) combined with reverse transcriptase activity. Editing cassettes are employed, comprising a gRNA and a repair template where the 3′ end of the repair template is protected from degradation.
Claims19
A CREATE fusion editing cassette for performing nucleic acid-guided nickase/reverse transcriptase fusion editing comprising from 3′ to 5′:
- an RNA repair template comprising:
- an RNA stabilization moiety;
- a linker region;
- a primer binding region capable of binding to a nicked target DNA;
- a nick-to-edit region;
- and
- a region of post-edit homology;
- a gRNA comprising:
- a guide sequence;
- and
- a scaffold region.
Dependencies are inferred from the claim text.