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In vivo genome editing in single mammalian brain neurons through CRISPR-Cas9 and cytosine base editorsopen access

Authors
Song, BeomjongKang, Chan YoungHan, Jun HeeKano, MasanobuKonnerth, ArthurBae, Sangsu
Issue Date
Jan-2021
Publisher
Elsevier
Keywords
CRISPR-Cas9; Base editors; In vivo targeted single-cell electroporation; Neuron; Brain
Citation
Computational and Structural Biotechnology Journal, v.19, pp 2477 - 2485
Pages
9
Indexed
SCIE
SCOPUS
Journal Title
Computational and Structural Biotechnology Journal
Volume
19
Start Page
2477
End Page
2485
URI
https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/194164
DOI
10.1016/j.csbj.2021.04.051
ISSN
2001-0370
Abstract
Gene manipulation is a useful approach for understanding functions of genes and is important for investigating basic mechanisms of brain function on the level of single neurons and circuits. Despite the development and the wide range of applications of CRISPR-Cas9 and base editors (BEs), their implementation for an analysis of individual neurons in vivo remained limited. In fact, conventional gene manipulations are generally achieved only on the population level. Here, we combined either CRISPR-Cas9 or BEs with the targeted single-cell electroporation technique as a proof-of-concept test for gene manipulation in single neurons in vivo. Our assay consisted of CRISPR-Cas9- or BEs-induced gene knockout in single Purkinje cells in the cerebellum. Our results demonstrate the feasibility of both gene editing and base editing in single cells in the intact brain, providing a tool through which molecular perturbations of individual neurons can be used for analysis of circuits and, ultimately, behaviors.
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