Institute of Hypoxia Medicine, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325015, China.
STAR Protoc. 2022 Apr 13;3(2):101296. doi: 10.1016/j.xpro.2022.101296. eCollection 2022 Jun 17.
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 and/or 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).
在人类多能干细胞（hPSCs）中，传统的基因过表达方法效率低，且通常很繁琐。在这里，我们提供了一种相对简单的方法，使用 Dox 诱导型 PiggyBac 转座子系统进行基因过表达。我们以 H1 胚胎干细胞（H1 ESCs）中过表达 和/或 为例，详细介绍了该方法的步骤。我们的方案可适用于各种 hPSCs 中的任何目的基因。如需了解本方案的详细使用和执行方法，请参考 Quan 等人（2021）。
