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Updated: Aug 27, 2025

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
Published on: May 5, 2023
Engineered CRISPR prime editors with compact, untethered reverse transcriptases
Julian Grünewald1,2,3,4,5,6,7, Bret R Miller8,9, Regan N Szalay8,9
1Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA. julian.grunewald@tum.de.
Abstract:
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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