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Virus-Mediated Genome Editing via Homology-Directed Repair in Mitotic and Postmitotic Cells in Mammalian Brain.

Authors
  • Nishiyama, Jun1
  • Mikuni, Takayasu2
  • Yasuda, Ryohei3
  • 1 Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA.
  • 2 Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA; Japan Science and Technology Agency, PRESTO, Kawaguchi, Saitama 332-0012, Japan. Electronic address: [email protected] , (Japan)
  • 3 Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA. Electronic address: [email protected]
Type
Published Article
Journal
Neuron
Publication Date
Nov 15, 2017
Volume
96
Issue
4
Identifiers
DOI: 10.1016/j.neuron.2017.10.004
PMID: 29056297
Source
Medline
Keywords
License
Unknown

Abstract

Precise genome editing via homology-directed repair (HDR) in targeted cells, particularly in vivo, provides an invaluable tool for biomedical research. However, HDR has been considered to be largely restricted to dividing cells, making it challenging to apply the technique in postmitotic neurons. Here we show that precise genome editing via HDR is possible in mature postmitotic neurons as well as mitotic cells in mice brain by combining CRISPR-Cas9-mediated DNA cleavage and the efficient delivery of donor template with adeno-associated virus (AAV). Using this strategy, we achieved efficient tagging of endogenous proteins in primary and organotypic cultures in vitro and developing, adult, aged, and pathological brains in vivo. Thus, AAV- and CRISPR-Cas9-mediated HDR will be broadly useful for precise genome editing in basic and translational neuroscience.

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