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Enabling large-scale genome editing at repetitive elements by reducing DNA nicking

Abstract : To extend the frontier of genome editing and enable editing of repetitive elements of mammalian genomes, we made use of a set of dead-Cas9 base editor (dBE) variants that allow editing at tens of thousands of loci per cell by overcoming the cell death associated with DNA double-strand breaks and single-strand breaks. We used a set of gRNAs targeting repetitive elements-ranging in target copy number from about 32 to 161 000 per cell. dBEs enabled survival after large-scale base editing, allowing targeted mutations at up to ∼13 200 and ∼12 200 loci in 293T and human induced pluripotent stem cells (hiP-SCs), respectively, three orders of magnitude greater than previously recorded. These dBEs can overcome current on-target mutation and toxicity barriers that prevent cell survival after large-scale genome engineering.
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https://hal.archives-ouvertes.fr/hal-03026263
Contributor : Hannu Myllykallio <>
Submitted on : Thursday, November 26, 2020 - 3:58:51 PM
Last modification on : Wednesday, December 16, 2020 - 4:06:00 AM

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Cory Smith, Oscar Castanon, Khaled Said, Verena Volf, Parastoo Khoshakhlagh, et al.. Enabling large-scale genome editing at repetitive elements by reducing DNA nicking. Nucleic Acids Research, Oxford University Press, 2020, 48 (9), pp.5183-5195. ⟨10.1093/nar/gkaa239⟩. ⟨hal-03026263⟩

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