Nanoparticle-mediated genome editing in single-cell embryos via peptide nucleic acids.

Autor: Putman R; Department of Biomedical Engineering Yale University New Haven Connecticut USA.; Department of Therapeutic Radiology Yale University New Haven Connecticut USA.; Duke University School of Medicine Durham North Carolina USA., Ricciardi AS; Department of Biomedical Engineering Yale University New Haven Connecticut USA.; Department of Therapeutic Radiology Yale University New Haven Connecticut USA.; Department of Surgery University of Pennsylvania Health Systems Philadelphia Pennsylvania USA., Carufe KEW; Department of Therapeutic Radiology Yale University New Haven Connecticut USA.; Department of Genetics Yale University New Haven Connecticut USA., Quijano E; Department of Biomedical Engineering Yale University New Haven Connecticut USA.; Department of Genetics Yale University New Haven Connecticut USA., Bahal R; Department of Therapeutic Radiology Yale University New Haven Connecticut USA.; Department of Pharmaceutical Sciences University of Connecticut Storrs Connecticut USA., Glazer PM; Department of Therapeutic Radiology Yale University New Haven Connecticut USA.; Department of Genetics Yale University New Haven Connecticut USA., Saltzman WM; Department of Biomedical Engineering Yale University New Haven Connecticut USA.; Department of Cellular & Molecular Physiology Yale University New Haven Connecticut USA.; Department of Chemical & Environmental Engineering Yale University New Haven Connecticut USA.
Jazyk: angličtina
Zdroj: Bioengineering & translational medicine [Bioeng Transl Med] 2022 Dec 02; Vol. 8 (3), pp. e10458. Date of Electronic Publication: 2022 Dec 02 (Print Publication: 2023).
DOI: 10.1002/btm2.10458
Abstrakt: Through preimplantation genetic diagnosis, genetic diseases can be detected during the early stages of embryogenesis, but effective treatments for many of these disorders are lacking. Gene editing could allow for correction of the underlying mutation during embryogenesis to prevent disease pathogenesis or even provide a cure. Here, we demonstrate that administration of peptide nucleic acids and single-stranded donor DNA oligonucleotides encapsulated in poly(lactic- co -glycolic acid) (PLGA) nanoparticles to single-cell embryos allows for editing of an eGFP-beta globin fusion transgene. Blastocysts from treated embryos exhibit high levels of editing (~94%), normal physiological development, normal morphology, and no detected off-target genomic effects. Treated embryos reimplanted to surrogate moms show normal growth without gross developmental abnormalities and with no identified off-target effects. Mice from reimplanted embryos consistently show editing, characterized by mosaicism across multiple organs with some organ biopsies showing up to 100% editing. This proof-of-concept work demonstrates for the first time the use of peptide nucleic acid (PNA)/DNA nanoparticles as a means to achieve embryonic gene editing.
Competing Interests: Raman Bahal, Peter M. Glazer, Elias Quijano, Adele S. Ricciardi, Rachael Putman, and W. Mark Saltzman are inventors on patent applications describing nanoparticle‐mediated delivery of triplex‐forming PNAs for gene editing. Raman Bahal is an inventor on a patent application regarding the synthesis of γPNAs. Rachael Putman, Adele S. Ricciardi, Peter M. Glazer, and W. Mark Saltzman are inventors on a patent application regarding embryonic gene editing in vitro.
(© 2022 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)
Databáze: MEDLINE
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