Production of TRPM4 knockout cell line using rat cardiomyocyte H9c2

Chen Wang, Masakazu Maeda, Jian Chen, Mengxue Wang, Keiji Naruse, Ken Takahashi

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


The method presented in this article are related to the research article entitled as “Role of the TRPM4 channel in mitochondrial function, calcium release, and ROS generation in oxidative stress" [1]. TRPM4, a non-selective monovalent cation channel, is not only involved in the generation of the action potential in cardiomyocytes, but also thought to be a key molecule in the development of the ischemia–reperfusion injury of the brain and the heart [2–5]. However, existing pharmacological inhibitors for the TRPM4 channel have problems of non-specificity [6]. This article describes methods used for targeted genomic deletion in the rat cardiomyocyte H9c2 using the CRISPR-Cas9 genome editing system in order to suppress TRPM4 protein expression. Confocal microscopy, flow cytometry, Sanger sequencing, and western blotting are performed to confirm vector transfection and the subsequent knockout of the TRPM4 protein. • These data provide information on the comprehensive analyses for knocking out the rat TRPM4 channel using CRISPR/Cas9. The analyses include confocal microscopy, flow cytometry, Sanger sequencing, and western blotting. • This dataset will benefit biological and medical researchers studying the function of TRPM4-expressing cells including neurons, cardiomyocytes, and vascular endothelial cells. It is also useful to study the involvement of the TRPM4 channel in pathological processes such as cardiac arrhythmia and ischemia–reperfusion injury. • The dataset can be used to guide the experiment of knocking out the TRPM4 gene and its subsequent application to the study of disease process caused by the gene.

Original languageEnglish
Article number101404
Publication statusPublished - Jan 2021


  • CRISPR knockout of rat TRPM4 gene
  • CRISPR/Cas9
  • Cardiomyocyte
  • Confocal microscopy
  • DNA sequencing
  • Flow cytometry
  • H9c2
  • TRPM4
  • Western blotting

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Medical Laboratory Technology


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