Protocol for cell preparation and gene delivery in HEK293T and C2C12 cells

Peng Jiang; Lejiao Ren; Li Zhi; Xinli Hu; Rui-Ping Xiao

doi:10.1016/j.xpro.2021.100497

Protocol for cell preparation and gene delivery in HEK293T and C2C12 cells

STAR Protoc. 2021 Jul 17;2(3):100497. doi: 10.1016/j.xpro.2021.100497. eCollection 2021 Sep 17.

Authors

Peng Jiang¹, Lejiao Ren², Li Zhi², Xinli Hu¹, Rui-Ping Xiao^{1

2}

Affiliations

¹ State Key Laboratory of Membrane Biology, Institute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China.
² Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.

Abstract

Exogenous overexpression of target genes in both general and specific cell types is important for mechanistic studies of gene function. Here, we provide a step-by-step protocol for cell culture, plasmid transfection in HEK293T, and adenoviral infection in C2C12 cells for gene overexpression in vitro, using MG53 as an example. This protocol enables sufficient and efficient gene expression for the downstream functional analysis. For complete details on the use and execution of this protocol, please refer to Jiang et al. (2021).

Keywords: Cell Biology; Cell culture; Cell-based Assays; Gene Expression; Molecular Biology.

Publication types

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

MeSH terms

Animals
Cell Differentiation
Cell Line
Gene Expression
Gene Transfer Techniques*
HEK293 Cells
Humans
Mice