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

eLife

Masahiro Shin¹  Feston Idrizi¹  Pengpeng Liu¹  Nathan D Lawson²  Yeonsoo Yoon³  Ping Xu³  Judith Gallant³  Jaime A Rivera-Pérez⁴  Xiao-Ou Zhang⁵  Krishna S Ghanta⁶  Zexiang Chen⁶  Gregoriy A Dokshin⁶  Alireza Edraki⁶  Pranathi M Krishnamurthy⁶  Ahmet Rasit Ozturk⁶  Aamir Mir⁶  Hassan Gneid⁷  Jonathan K Watts⁸  Erik J Sontheimer⁹  Craig C Mello^1,10 

[1] Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States;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;Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States;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;Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, United States;RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States;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;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;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;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;

关键词: CRISPR;    HDR;    modified donors;    chemical modifications;    genome editing;    Human;    Mouse;    C. elegans;    Zebrafish;   

DOI  :  10.7554/eLife.72216

来源: eLife Sciences Publications, Ltd

【 摘 要 】

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.

【 授权许可】

CC BY