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

Jiwen Yang; Minjie Hu; Yongyu Wang

doi:10.1016/j.xpro.2022.101296

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. eCollection 2022 Jun 17.

Authors

Jiwen Yang¹, Minjie Hu¹, Yongyu Wang¹

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA Transposable Elements* / genetics
Humans
Pluripotent Stem Cells*
Transgenes / genetics

Substances

DNA Transposable Elements