PubMed ID: 42595755
Author(s): Saxena S, Kabra M, Abdeen AA, Tabima DM, Sinha D, Rawding PA, Zhu M, Xie R, Kulkarni T, Hanstad GM, Fernandez Zepeda MA, Gamm DM, Pattnaik BR, Gong S, Saha K. Genome-wide CRISPR screening identifies cellular factors controlling nonviral genome editing efficiency. Nat Commun. 2026 Aug 13;17(1):8086. doi: 10.1038/s41467-026-76350-5. PMID: 42595755.PMID 42595755
Journal: Nature Communications 17(1)
To systematically map cellular factors constraining nonviral genome editing, influencing uptake and intracellular trafficking, we develop a genome-wide CRISPR screening platform linking perturbation of 19,114 genes to editing outcomes in human cells. We identify six negative regulators of delivery whose depletion increases editing efficiency by up to six-fold across diverse payloads, loci, and cell types. We test the top two factors, GJB2 and BET1L, in two distinct human models: correction of a pathogenic adenine base mutation in KCNJ13 and introduction of a cytosine base mutation in the GABAA receptor gene. Depletion of either improves base-editing outcomes by 6-fold, potentially through effects on delivery. In a patient-derived model of retinal channelopathy, knockdown of either gene improves lipid nanoparticle base editing efficiency by over 3.5-fold. This enables functional restoration of Kir7.1 ion channels in a subset of edited cells, highlighting cellular barriers as actionable targets to enhance the potency of genetic therapies.