Protocol to perform multiplexed assays of variant effect using curated loci prime editing.

ABSTRACT: Multiplexed assays of variant effect (MAVEs) perform simultaneous characterization of many variants. Here, we present a protocol to perform MAVEs using curated loci prime editing (cliPE), an accessible experimental pipeline that enables prime editing of a target gene. We describe steps for designing prime editing reagents, screening for genome editing efficiency, selecting a pool of cells edited to harbor different genetic variants, and sequencing. Lastly, we detail procedures for performing enrichment analysis to identify variants with normal or aberrant activity.

SUBMITTER: Biar CG 

PROVIDER: S-EPMC12159904 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature