Protocol for inducible piggyBac transposon system for efficient gene overexpression in human pluripotent stem cells

  • STAR Protoc. 2022 Apr 13;3(2):101296. doi: 10.1016/j.xpro.2022.101296.
Jiwen Yang  1 ,  Minjie Hu  1 ,  Yongyu Wang  1
Affiliations
  • 1. Institute of Hypoxia Medicine, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325015, China.
PMID: 35496784 DOI: 10.1016/j.xpro.2022.101296
Abstract

In human pluripotent stem cells (hPSCs), traditional approaches for gene overexpression have low efficiency and are often laborious. Here, we provide a relatively simple protocol for gene overexpression with the Dox-inducible PiggyBac transposon system. We detail the steps for overexpression of FLI1 and/or YAP in H1 embryonic stem cells (H1 ESCs) as an example. Our protocol can be applied to any gene of interest in a variety of hPSCs. For complete details on the use and execution of this protocol, please refer to Quan et al. (2021).

Keywords
Cell Biology; Cell Differentiation; Developmental biology; Molecular Biology; Stem Cells.
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