eLife (Oct 2021)
5′-Modifications improve potency and efficacy of DNA donors for precision genome editing
- Krishna S Ghanta,
- Zexiang Chen,
- Aamir Mir,
- Gregoriy A Dokshin,
- Pranathi M Krishnamurthy,
- Yeonsoo Yoon,
- Judith Gallant,
- Ping Xu,
- Xiao-Ou Zhang,
- Ahmet Rasit Ozturk,
- Masahiro Shin,
- Feston Idrizi,
- Pengpeng Liu,
- Hassan Gneid,
- Alireza Edraki,
- Nathan D Lawson,
- Jaime A Rivera-Pérez,
- Erik J Sontheimer,
- Jonathan K Watts,
- Craig C Mello
Affiliations
- Krishna S Ghanta
- ORCiD
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Zexiang Chen
- ORCiD
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Aamir Mir
- ORCiD
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Gregoriy A Dokshin
- ORCiD
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Pranathi M Krishnamurthy
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Yeonsoo Yoon
- ORCiD
- Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States
- Judith Gallant
- Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States
- Ping Xu
- Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, Worcester, United States
- Xiao-Ou Zhang
- Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, United States
- Ahmet Rasit Ozturk
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Masahiro Shin
- ORCiD
- Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States
- Feston Idrizi
- ORCiD
- Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States
- Pengpeng Liu
- Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, United States
- Hassan Gneid
- ORCiD
- 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
- Alireza Edraki
- RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States
- Nathan D Lawson
- ORCiD
- 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
- Jaime A Rivera-Pérez
- 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
- Erik J Sontheimer
- ORCiD
- 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
- Jonathan K Watts
- ORCiD
- 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
- Craig C Mello
- ORCiD
- 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
- DOI
- https://doi.org/10.7554/eLife.72216
- Journal volume & issue
-
Vol. 10
Abstract
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.
Keywords
