Work

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

Year: 2025

Type: article

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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103851

Authors: Carina G. Biar, Nicholas Bodkin, Gemma L. Carvill, Jeffrey D. Calhoun

Institutions: University of Washington, Northwestern University, Northwestern Medicine

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: American Epilepsy Society, Northwestern University, National Institutes of Health, National Cancer Institute