Enhancement of CRISPR gene editing or target destruction by co-expression of heterologous DNA repair protein
AssigneeThe Rockefeller University
InventorsLuciano Marraffini, Jon McGinn, Josh Modell, Dominik Paquet
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Abstract
Provided are compositions, methods, and kits for improving CRISPR-based editing of DNA targets by a CRISPR-associated (Cas) enzyme. The improvement is made by combining the Cas enzyme and a CRISPR targeting RNA a heterologous DNA repair enzyme that is at least one of RecBCD, AddAB, or AdnAB. The heterologous DNA repair enzyme may have inactivated nuclease activity. The method can include using a DNA repair template to introduce one or more changes into the edited DNA. Cells that contain components of the improved CRISPR systems are included, as are kits for making such cells.
Claims8
A method comprising:
- providing a cell comprising one or more DNA elements that encode:
- i) a CRISPR-associated Cas9 enzyme;
- ii) a heterologous DNA repair enzyme that is at least one of RecBCD, AddAB, or AdnAB, wherein optionally the RecBCD, AddAB, or AdnAB in the cell has inactivated nuclease activity, and
- iii) a targeting RNA
- wherein the Cas9 enzyme and at least one of the heterologous RecBCD, AddAB, or AdnAB are expressed, and wherein editing of DNA in the cell that is targeted by the targeting RNA occurs.
Dependencies are inferred from the claim text.