Bibliographic Details
| Title: |
Precise genomic deletions using paired prime editing. |
| Authors: |
Choi, Junhong, Chen, Wei, Suiter, Chase C., Lee, Choli, Chardon, Florence M., Yang, Wei, Leith, Anh, Daza, Riza M., Martin, Beth, Shendure, Jay |
| Source: |
Nature Biotechnology; Feb2022, Vol. 40 Issue 2, p218-226, 9p |
| Abstract: |
Current methods to delete genomic sequences are based on clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 and pairs of single-guide RNAs (sgRNAs), but can be inefficient and imprecise, with errors including small indels as well as unintended large deletions and more complex rearrangements. In the present study, we describe a prime editing-based method, PRIME-Del, which induces a deletion using a pair of prime editing sgRNAs (pegRNAs) that target opposite DNA strands, programming not only the sites that are nicked but also the outcome of the repair. PRIME-Del achieves markedly higher precision than CRISPR–Cas9 and sgRNA pairs in programming deletions up to 10 kb, with 1–30% editing efficiency. PRIME-Del can also be used to couple genomic deletions with short insertions, enabling deletions with junctions that do not fall at protospacer-adjacent motif sites. Finally, extended expression of prime editing components can substantially enhance efficiency without compromising precision. We anticipate that PRIME-Del will be broadly useful for precise, flexible programming of genomic deletions, epitope tagging and, potentially, programming genomic rearrangements. Paired prime editing is used to precisely delete genomic sequences up to 10 kb. [ABSTRACT FROM AUTHOR] |
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| Database: |
Complementary Index |
