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人类神经干细胞衍生神经元中转录和表观遗传因子的单分子成像方案。

Protocol for single-molecule imaging of transcription and epigenetic factors in human neural stem cell-derived neurons.

作者信息

Atsumi Yuri, Yamamoto Nobuhiko, Sugo Noriyuki

机构信息

Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.

Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan; Institute of Neurological and Psychiatric Disorders, Shenzhen Bay Laboratory, Shenzhen, Guangdong 518132, China.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103432. doi: 10.1016/j.xpro.2024.103432. Epub 2024 Nov 1.

Abstract

Single-molecule imaging (SMI) is a powerful approach to quantify the spatiotemporal dynamics of transcription in living cells. Here, we describe a protocol of SMI for transcription and epigenetic factors in human cortical neurons derived from embryonic stem cells or induced pluripotent stem cells. Specifically, we detail the procedures for neural stem cell culture, gene transfer, microscopy, and data analysis. This protocol can be applied to the study of transcription dynamics in response to various cellular stimuli. For complete details on the use and execution of this protocol, please refer to Atsumi et al..

摘要

单分子成像(SMI)是一种用于量化活细胞中转录时空动态的强大方法。在这里,我们描述了一种针对源自胚胎干细胞或诱导多能干细胞的人类皮质神经元中转录和表观遗传因子的SMI方案。具体而言,我们详细介绍了神经干细胞培养、基因转移、显微镜检查和数据分析的程序。该方案可应用于研究对各种细胞刺激作出反应的转录动态。有关该方案使用和执行的完整详细信息,请参考敦美等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bed7/11565389/f9ee3b62db9c/fx1.jpg

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