A protocol to investigate the effects of lncRNAs on in vivo protein-protein interactions using proximity ligation assay

Ling Zhang; Mengfan He; Peizhen Wang; Jianfeng Yu; Dawei Li

doi:10.1016/j.xpro.2023.102757

A protocol to investigate the effects of lncRNAs on in vivo protein-protein interactions using proximity ligation assay

STAR Protoc. 2023 Dec 15;4(4):102757. doi: 10.1016/j.xpro.2023.102757. Epub 2023 Dec 2.

Authors

Ling Zhang¹, Mengfan He¹, Peizhen Wang¹, Jianfeng Yu², Dawei Li³

Affiliations

¹ Center for Translational Medicine, The Affiliated Zhangjiagang Hospital of Soochow University, Suzhou Medical College of Soochow University, 68 Jiyang West Road, Suzhou 215600, China.
² Center for Translational Medicine, The Affiliated Zhangjiagang Hospital of Soochow University, Suzhou Medical College of Soochow University, 68 Jiyang West Road, Suzhou 215600, China; Department of Life Science and Technology, Changshu Institute of Technology, 99 South Third Ring Road, Suzhou 215500, China.
³ Center for Translational Medicine, The Affiliated Zhangjiagang Hospital of Soochow University, Suzhou Medical College of Soochow University, 68 Jiyang West Road, Suzhou 215600, China. Electronic address: daweili@suda.edu.cn.

Abstract

A large variety of cellular signals are triggered and transmitted by protein-protein interactions (PPIs). Long noncoding RNAs regulate PPIs by enhancing or destabilizing these interactions. Here, we use the proximity ligation assay technique to determine PPIs between p53 and SET regulated by long intergenic noncoding RNA 324 (LINC00324). We detail procedures for establishing LINC00324 knockdown and overexpression U2OS and HepG2 cells followed by in situ PLA protocol. This approach has many potential applications for the study of cellular factors that regulate PPIs. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2023).¹.

Keywords: Cancer; Cell Biology; Microscopy; Molecular/Chemical Probes; Protein Biochemistry.

MeSH terms

RNA, Long Noncoding* / genetics
Signal Transduction

Substances

RNA, Long Noncoding