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.
You may also like
Hitchhiking through the nerve cell
Getting in the Groove: Why samba makes everyone want...
Estimating the pace of change
April 26, 2022Max Planck Institute for Biological Cybernetics
Out of rhythm: Compromised precision of theta-gamma...
April 26, 2022Max Planck Institute for Human Development
Near-natural, fractal architecture promotes well-being
April 26, 2022Max Planck Institute for Biological Cybernetics
Decoding cognition from spontaneous neural activity
April 26, 2022Max Planck Institute for Human Development