Curated loci prime editing (cliPE) for accessible multiplexed assays of variant effect (MAVEs).

ArXiv Pub Date : 2025-01-08
Carina G Biar, Nicholas Bodkin, Gemma L Carvill, Jeffrey D Calhoun
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Abstract

Multiplexed assays of variant effect (MAVEs) perform simultaneous characterization of many variants. Prime editing has been recently adopted for introducing many variants in their native genomic contexts. However, robust protocols and standards are limited, preventing widespread uptake. Herein, we describe curated loci prime editing (cliPE) which is an accessible, low-cost experimental pipeline to perform MAVEs using prime editing of a target gene, as well as a companion Shiny app (pegRNA Designer) to rapidly and easily design user-specific MAVE libraries.

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变异效应多重检测(MAVEs)可同时鉴定许多变异。Prime editing 最近已被采用,用于在原生基因组环境中引入许多变体。然而,稳健的协议和标准有限,阻碍了其广泛应用。在这里,我们将介绍一种可利用的、低成本的实验管道--cliPE(curated loci prime editing),该管道可利用目标基因的质粒编辑来执行 MAVEs,我们还将介绍一种配套的 Shiny 应用程序(pegRNA Designer),该程序可快速、轻松地设计用户特异的 MAVE 文库。
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