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Engineered CRISPR prime editors with compact, untethered reverse transcriptases.

Julian GrünewaldBret R MillerRegan N SzalayPeter K CabeceirasChristopher J WoodillaEliza Jane B HoltzKarl PetriJ Keith Joung
Published in: Nature biotechnology (2022)
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.
Keyphrases
  • crispr cas
  • genome editing
  • genome wide
  • bone marrow
  • dna methylation
  • gene expression
  • candida albicans
  • low cost
  • cystic fibrosis
  • pseudomonas aeruginosa