Virus-Mediated Genome Editing via Homology-Directed Repair in Mitotic and Postmitotic Cells in Mammalian Brain
Open accessAbstract
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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Although some homology-directed repair has been observed in non-dividing cells such as neurons, other DNA repair pathways than homologous recombination are more active in these long-lived postmitotic cells.
“Although a certain level of homology-directed repair has been observed in non-dividing cells, such as neurons [52], other DNA repair pathways than homologous recombination are more active in these long-lived postmitotic cells [53].”
From Circumventing the packaging limit of AAV-mediated gene replacement therapy for neurological disorders (2022) · cites this paper as [52]
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“rons [52], other DNA repair pathways than homologous recombination are more active in these long-lived postmitotic cells [53].”
Background
- Authors
- Nishiyama, J., Mikuni, T., & Yasuda, R.
- Affiliations
- Max Planck Florida Institute for Neuroscience (US)
- Japan Science and Technology Agency (JP)
- Corresponding author
- Mikuni, Takayasu
- Yasuda, Ryohei
- Venue
- Neuron
- Publisher
- Elsevier BV
- Published
- 2017-11
- Volume
- 96
- Issue
- 4
- Pages
- 755-768.e5
- ISSN
- 0896-6273 (ISSN-L)
- PMC
- PMC5691606
- OpenAlex
- W2765658539
- Cited by
- 215
- References
- 11
- Citations by year
- 2026: 16 · 2025: 18 · 2024: 17 · 2023: 23 · 2022: 20 · 2021: 30 · 2020: 41 · 2019: 27 · 2018: 22 · 2017: 1
- FWCI
- 10.17
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 379 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Neuron (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Funders
- National Institute of Health (R01MH080047, DP1NS096787)
- Human Frontier Science Program (LT000974/2014-L)
- JST/PRESTO, Japan (JPMJPR16F9)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- homology-directed repair, CRISPR/Cas9, adeno-associated virus, in vivo gene editing, postmitotic neurons, mitotic cells, brain, protein labeling, organotypic brain cultures, brain aging
- MeSH terms
- Gene Editing, Dependovirus, Animals, Brain, Mitosis, Neurons, Mice, Rats, Recombinational DNA Repair, CRISPR-Cas Systems
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