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Engineered CRISPR prime editors with compact, untethered reverse transcriptases

Nat Biotechnol. 2022-09; 
Julian Grünewald, Bret R Miller, Regan N Szalay, Peter K Cabeceiras, Christopher J Woodilla, Eliza Jane B Holtz, Karl Petri, J Keith Joung
Products/Services Used Details Operation
Plasmid DNA Preparation … encoding alternative RTs were purchased from IDT as synthetic double-stranded DNA (dsDNA) products (IDT gblocks) with codon optimization for expression in human cells (GenScript Get A Quote

摘要

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.

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