Journal
NATURE BIOTECHNOLOGY
Volume 41, Issue 3, Pages 337-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41587-022-01473-1
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The CRISPR prime editor PE2, composed of nSpCas9 and MMLV-RT, functions efficiently in human cells. The Split-PE system allows for the identification and engineering of more compact prime editor architectures that broaden the types of RTs used for prime editing. A split prime editor architecture facilitates screening and engineering of improved variants.
The CRISPR prime editor PE2 consists of a Streptococcus pyogenes Cas9 nickase (nSpCas9) fused at its C-terminus to a Moloney murine leukemia virus reverse transcriptase (MMLV-RT). Here we show that separated nSpCas9 and MMLV-RT proteins function as efficiently as intact PE2 in human cells. We use this Split-PE system to rapidly identify and engineer more compact prime editor architectures that also broaden the types of RTs used for prime editing. A split prime editor architecture facilitates screening and engineering of improved variants.
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