1. Academic Validation
  2. Protocol for inducible piggyBac transposon system for efficient gene overexpression in human pluripotent stem cells

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

Affiliation

  • 1 Institute of Hypoxia Medicine, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325015, China.
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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