2. Academic Validation
  3. Workflow for Performing Genetic Manipulation in Human Trophoblast Stem Cells Using CRISPR/Cas9 Technology

Workflow for Performing Genetic Manipulation in Human Trophoblast Stem Cells Using CRISPR/Cas9 Technology

  • Methods Mol Biol. 2022 Dec 15. doi: 10.1007/7651_2022_464.
Hao Wu 1 2 3 4,  Yue Wang 1 2 3 4,  Hongmei Wang 5 6 7 8
Affiliations

Affiliations

  • 1 State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • 2 Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China.
  • 3 University of Chinese Academy of Sciences, Beijing, China.
  • 4 Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China.
  • 5 State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China. wanghm@ioz.ac.cn.
  • 6 Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China. wanghm@ioz.ac.cn.
  • 7 University of Chinese Academy of Sciences, Beijing, China. wanghm@ioz.ac.cn.
  • 8 Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China. wanghm@ioz.ac.cn.
PMID: 36515893 DOI: 10.1007/7651_2022_464
Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-based tool kits greatly facilitate the investigation of gene function, especially gene editing and transcriptional regulation technology. Human trophoblast stem cell (hTSC) is a promising cell model for simulating the development and function of human placental trophoblast cells. Here, we provide an optimized workflow for efficient genome editing and transcriptional activation in hTSCs using a lentivirus-dependent method. We also present methods to evaluate the gene targeted mutations and transcriptional activation.

Keywords

CRISPR/Cas9; CRISPR/dCas9-VP64; Human trophoblast stem cell.

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