5'-Modifications improve potency and efficacy of DNA donors for precision genome editing.

Autor:	Ghanta KS; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Chen Z; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Mir A; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Dokshin GA; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Krishnamurthy PM; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Yoon Y; Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States., Gallant J; Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States., Xu P; Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States., Zhang XO; Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, United States., Ozturk AR; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Shin M; Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States., Idrizi F; Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States., Liu P; Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States., Gneid H; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States.; Department of Biochemistry and Molecular Biotechnology, University of Massachusetts Medical School, Worcester, United States., Edraki A; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States., Lawson ND; Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States.; Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, United States.; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, United States., Rivera-Pérez JA; Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States.; Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, United States., Sontheimer EJ; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States.; Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, United States.; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, United States., Watts JK; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States.; Department of Biochemistry and Molecular Biotechnology, University of Massachusetts Medical School, Worcester, United States.; Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, United States., Mello CC; RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States.; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, United States.; Howard Hughes Medical Institute, University of Massachusetts Medical School, Worcester, United States.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2021 Oct 19; Vol. 10. Date of Electronic Publication: 2021 Oct 19.
DOI:	10.7554/eLife.72216
Abstrakt:	Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as a therapeutic approach to correct mutations that cause disease. In its most precise form, genome editing can use cellular homology-directed repair (HDR) pathways to insert information from an exogenously supplied DNA-repair template (donor) directly into a targeted genomic location. Unfortunately, particularly for long insertions, toxicity and delivery considerations associated with repair template DNA can limit HDR efficacy. Here, we explore chemical modifications to both double-stranded and single-stranded DNA-repair templates. We describe 5'-terminal modifications, including in its simplest form the incorporation of triethylene glycol (TEG) moieties, that consistently increase the frequency of precision editing in the germlines of three animal models ( Caenorhabditis elegans , zebrafish, mice) and in cultured human cells. Competing Interests: KG, GD, HG, ES, JW Co-inventor on patent applications related to this work (Application number: 16/384, 612 ), ZC, PK, YY, JG, PX, XZ, AO, MS, FI, PL, AE, NL, JR No competing interests declared, AM Co-inventor on patent application related to this work (Application number: 16/384, 612 ), CM The authors (K.S.G, A.M, G.A.D, H.G, J.K.W, E.J.S and C.C.M) have a patent application pending related to the findings described (Application number: 16/384, 612 ). Craig C. Mello is a co-founder and Scientific Advisory Board member of CRISPR Therapeutics, and Erik J. Sontheimer is a cofounder and Scientific Advisory Board member of Intellia herapeutics. (© 2021, Ghanta et al.)
Databáze:	MEDLINE
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