5' modifications improve potency and efficacy of DNA donors for precision genome editing
- University of Massachusetts Medical School, United States
- UMass medical School, United States
- UMass Medical School, United States
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.
Data availability
All the sequencing data will be deposited to Dyrad and can be accessed at https://doi.org/10.5061/dryad.f7m0cfxwr
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Data from: 5′ Modifications Improve Potency and Efficacy of DNA Donors for Precision Genome EditingDryad Digital Repository, doi.10.5061/dryad.f7m0cfxwr.
Article and author information
Author details
Funding
Howard Hughes Medical Institute
- Craig C Mello
Office of Extramural Research, National Institutes of Health (R37 GM058800-23)
- Craig C Mello
National Center for Advancing Translational Sciences (UG3 TR002668)
- Erik Sontheimer
- Jonathan K Watts
National Center for Advancing Translational Sciences (UG3 TR002668)
- Erik Sontheimer
National Heart, Lung, and Blood Institute (R35 HL140017)
- Nathan D Lawson
NIH Office of the Director (R21 OD030004)
- Nathan D Lawson
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: Fish were maintained in accordance with the protocols set by the University of Massachusetts Medical School Institutional Animal Care and Use Committee. All the mouse experiments were conducted according the UMMS Institute Animal Care and Use Committee (IACUC).
Copyright
© 2021, Ghanta et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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5' modifications improve potency and efficacy of DNA donors for precision genome editing
eLife 10:e72216.
https://doi.org/10.7554/eLife.72216
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