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
  • 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. [email protected].
  • 6. Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China. [email protected].
  • 7. University of Chinese Academy of Sciences, Beijing, China. [email protected].
  • 8. Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China. [email protected].
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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