@article{oai:soar-ir.repo.nii.ac.jp:00020253,
 author = {Uemura, Takeshi and Mori, Takuma and Kurihara, Taiga and Kawase, Shiori and Koike, Rie and Satoga, Michiru and Cao, Xueshan and Li, Xue and Yanagawa, Toru and Sakurai, Takayuki and Shindo, Takayuki and Tabuchi, Katsuhiko},
 journal = {SCIENTIFIC REPORTS},
 month = {Oct},
 note = {Genome editing is a powerful technique for studying gene functions. CRISPR/Cas9-mediated gene knock-in has recently been applied to various cells and organisms. Here, we successfully knocked in an EGFP coding sequence at the site immediately after the first ATG codon of the β-actin gene in neurons in the brain by the combined use of the CRISPR/Cas9 system and in utero electroporation technique, resulting in the expression of the EGFP-tagged β-actin protein in cortical layer 2/3 pyramidal neurons. We detected EGFP fluorescence signals in the soma and neurites of EGFP knock-in neurons. These signals were particularly abundant in the head of dendritic spines, corresponding to the localization of the endogenous β-actin protein. EGFP knock-in neurons showed no detectable changes in spine density and basic electrophysiological properties. In contrast, exogenously overexpressed EGFP-β-actin showed increased spine density and EPSC frequency, and changed resting membrane potential. Thus, our technique provides a potential tool to elucidate the localization of various endogenous proteins in neurons by epitope tagging without altering neuronal and synaptic functions. This technique can be also useful for introducing a specific mutation into genes to study the function of proteins and genomic elements in brain neurons., Article, SCIENTIFIC REPORTS.6:35861(2016)},
 title = {Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques},
 volume = {6},
 year = {2016}
}