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

被引：0
|
作者：
Atsumi, Yuri [1 ]
Yamamoto, Nobuhiko [1 ,2 ]
Sugo, Noriyuki [1 ]
机构：
[1] Osaka Univ, Grad Sch Frontier Biosci, Suita, Osaka 5650871, Japan
[2] Shenzhen Bay Lab, Inst Neurol & Psychiat Disorders, Shenzhen 518132, Guangdong, Peoples R China
来源：
STAR PROTOCOLS | 2024年 / 5卷 / 04期
关键词：
CORTICAL-NEURONS; DYNAMICS; BINDING; PROTEIN;
D O I：
10.1016/j.xpro.2024.103432
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.1
引用
收藏
页数：18
相关论文
共 7 条
  • [1] Live cell single-molecule imaging to study DNA repair in human cells
    Heyza, Joshua R.
    Mikhova, Mariia
    Schmidt, Jens C.
    DNA REPAIR, 2023, 129
  • [2] Single-cell analysis of diversity in human stem cell-derived neurons
    Harbom, Lise J.
    Michel, Nadine
    McConnell, Michael J.
    CELL AND TISSUE RESEARCH, 2018, 371 (01) : 171 - 179
  • [3] Single-molecule imaging of the transcription factor SRF reveals prolonged chromatin-binding kinetics upon cell stimulation
    Hipp, Lisa
    Beer, Judith
    Kuchler, Oliver
    Reisser, Matthias
    Sinske, Daniela
    Michaelis, Jens
    Gebhardt, J. Christof M.
    Knoell, Bernd
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2019, 116 (03) : 880 - 889
  • [4] Human embryonic stem cell-derived neurons adopt and regulate the activity of an established neural network
    Weick, Jason P.
    Liu, Yan
    Zhang, Su-Chun
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2011, 108 (50) : 20189 - 20194
  • [5] Injured inflammatory environment overrides the TET2 shaped epigenetic landscape of pluripotent stem cell derived human neural stem cells
    Kamei, Noriko
    Day, Kenneth
    Guo, Wei
    Haus, Daniel L.
    Nguyen, Hal X.
    Scarfone, Vanessa M.
    Booher, Keith
    Jia, Xi-Yu
    Cummings, Brian J.
    Anderson, Aileen J.
    SCIENTIFIC REPORTS, 2024, 14 (01):
  • [6] Comparison of the Gene Expression Profiles of Human Fetal Cortical Astrocytes with Pluripotent Stem Cell Derived Neural Stem Cells Identifies Human Astrocyte Markers and Signaling Pathways and Transcription Factors Active in Human Astrocytes
    Malik, Nasir
    Wang, Xiantao
    Shah, Sonia
    Efthymiou, Anastasia G.
    Yan, Bin
    Heman-Ackah, Sabrina
    Zhan, Ming
    Rao, Mahendra
    PLOS ONE, 2014, 9 (05):
  • [7] Alpha7 nicotinic acetylcholine receptors and neural network synaptic transmission in human induced pluripotent stem cell-derived neurons
    Larsen, Hjalte M.
    Hansen, Susanne K.
    Mikkelsen, Jens D.
    Hyttel, Poul
    Stummann, Tina C.
    STEM CELL RESEARCH, 2019, 41
1