Search-and-replace genome editing without double-strand breaks or donor DNA

Search-and-replace genome editing without double-strand breaks or donor DNA

2019 December ; 576(7785): 149–157. doi:10.1038/s41586-019-1711-4 | Andrew V. Anzalone, Peyton B. Randolph, Jessie R. Davis, Alexander A. Sousa, Luke W. Koblan, Jonathan M. Levy, Peter J. Chen, Christopher Wilson, Gregory A. Newby, Aditya Raguram, David R. Liu
The authors describe a new genome editing method called prime editing, which allows for precise and versatile genetic modifications without the need for double-strand breaks or donor DNA templates. Prime editing uses a catalytically impaired Cas9 fused to an engineered reverse transcriptase (RT) and a prime editing guide RNA (pegRNA) to directly write new genetic information into a specified DNA site. The method can perform various types of edits, including insertions, deletions, and all 12 types of point mutations, with high efficiency and few byproducts. Prime editing was tested in human cells and mouse neurons, showing higher or similar efficiency and fewer byproducts compared to homology-directed repair and base editing, and significantly lower off-target editing than Cas9 nuclease. The authors demonstrate the potential of prime editing to correct genetic diseases, such as sickle cell disease and Tay-Sachs disease, and to install protective mutations. Prime editing offers a new "search-and-replace" capability that expands the scope of genome editing, potentially correcting up to 89% of known genetic variants associated with human diseases.The authors describe a new genome editing method called prime editing, which allows for precise and versatile genetic modifications without the need for double-strand breaks or donor DNA templates. Prime editing uses a catalytically impaired Cas9 fused to an engineered reverse transcriptase (RT) and a prime editing guide RNA (pegRNA) to directly write new genetic information into a specified DNA site. The method can perform various types of edits, including insertions, deletions, and all 12 types of point mutations, with high efficiency and few byproducts. Prime editing was tested in human cells and mouse neurons, showing higher or similar efficiency and fewer byproducts compared to homology-directed repair and base editing, and significantly lower off-target editing than Cas9 nuclease. The authors demonstrate the potential of prime editing to correct genetic diseases, such as sickle cell disease and Tay-Sachs disease, and to install protective mutations. Prime editing offers a new "search-and-replace" capability that expands the scope of genome editing, potentially correcting up to 89% of known genetic variants associated with human diseases.
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[slides and audio] Search-and-replace genome editing without double-strand breaks or donor DNA