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Article

A rapid and robust protocol for generating loss-of-function alleles in pluripotent stem cells.

STAR protocols - 19 Sept 2025

Baljinnyam Erdene, Grisanti Laura, Tattari Shalini, Pedicone Chiara, Shewale Bhavana, Dubois Nicole, Goate Alison M, Kerr William G, Marro Samuele G

Abstract excerpt

The generation of loss-of-function alleles in human pluripotent stem cells (hPSCs) is used to interrogate gene function and validate reagents; however, identifying clones harboring true loss-of-function alleles remains inefficient. To address this, we present BOLT (barcoded oligos for loss-of-function targeting), a streamlined protocol that simplifies the screening process, facilitating rapid validation of...

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Identifiers

PMID: 40616838
DOI: 10.1016/j.xpro.2025.103916
PMCID: PMC12272864

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