Precise genome editing via homology-directed repair (HDR) in targeted cells, particularlyin 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 culturesin vitroand developing, adult, aged, and pathological brainsin vivo. Thus, AAV- and CRISPR-Cas9-mediated HDR will be broadly useful for precise genome editing in basic and translational neuroscience.

中文翻译：

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病毒介导的基因组编辑，通过同源性定向修复哺乳动物脑中的有丝分裂和有丝分裂后的细胞

在靶细胞中，特别是在体内，通过同源直接修复（HDR）进行精确的基因组编辑，为生物医学研究提供了宝贵的工具。然而，HDR被认为在很大程度上限于分裂细胞，这使其在有丝分裂后的神经元中应用该技术具有挑战性。在这里我们显示，通过结合CRISPR-Cas9介导的DNA裂解和腺相关病毒（AAV）高效递送供体模板，可以通过HDR在成熟的有丝分裂后神经元以及小鼠脑中的有丝分裂细胞中进行精确的基因组编辑。使用此策略，我们实现了体外原代和器官型培养以及体内成年，成年，老年和病理性脑的有效内源蛋白标记。因此，