Compositions, methods, and systems for genome editing technology
AssigneeNorth Carolina State University
InventorsChase Lawrence Beisel, Scott Collins
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Abstract
The present disclosure provides compositions, methods, and systems related to genome editing technology. In particular, the present disclosure provides a novel CRISPR-based genome editing technology that involves the generation of abasic sites to facilitate genetic recombination, without the need for breaks in the DNA. The compositions, methods, and systems described herein address many of the drawbacks of currently available approaches, including off-target effects and cellular toxicity.
Claims38
A composition for targeted editing of a nucleic acid, the composition comprising:
- a) a gap editor complex comprising a DNA-recognition domain and a DNA-modifying domain, wherein the gap editor complex induces formation of at least one abasic site in a target sequence of a double-stranded DNA molecule;
- b) at least one guide RNA molecule;
- and
- c) a donor nucleic acid template, wherein the presence of the donor nucleic acid template facilitates homology-directed recombination, and wherein the donor nucleic acid template or a fragment thereof is recombined into the double-stranded DNA molecule.
A kit for targeted editing of a nucleic acid, the kit comprising:
- a) a gap editor complex comprising a DNA-recognition domain and a DNA-modifying domain, wherein the gap editor complex induces formation of at least one abasic site in a target sequence of a double-stranded DNA molecule;
- b) at least one guide RNA molecule;
- and
- c) a donor nucleic acid template, wherein the presence of the donor nucleic acid template facilitates homology-directed recombination, and wherein the donor nucleic acid template or a fragment thereof is recombined into the double-stranded DNA molecule.
A system for targeted editing of a nucleic acid, the system comprising:
- a) a gap editor complex comprising a DNA-recognition domain and a DNA-modifying domain, wherein the gap editor complex induces formation of at least one abasic site in a target sequence of a double-stranded DNA molecule;
- b) at least one guide RNA molecule;
- and
- c) a donor nucleic acid template, wherein the presence of the donor nucleic acid template facilitates homology-directed recombination, and wherein the donor nucleic acid template or a fragment thereof is recombined into the double-stranded DNA molecule.
Dependencies are inferred from the claim text.