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Title: Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture
Authors: Bai, Qianfan
Bai, Zhiqiang
Xu, Shaohai
Sun, Lei
Keywords: Molecular Biology
RNA-binding protein
Issue Date: 2016
Source: Bai, Q., Bai, Z., Xu, S., & Sun, L. (2016). Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture. Journal of Visualized Experiments, 113, e54207-.
Series/Report no.: Journal of Visualized Experiments
Abstract: RNA-binding proteins (RBPs) are emerging as a regulatory layer in the development and function of adipose. RBPs play a key role in the gene expression regulation at posttranscriptional levels by affecting the stability and translational efficiency of target mRNAs. RNA pull-down technique has been widely used to study RNA-protein interaction, which is necessary to elucidate the mechanism underlying RBPs' as well as long non-coding RNAs' (lncRNAs) function. However, the high lipid abundance in adipocytes poses a technical challenge in conducting this experiment. Here a detailed RNA pull-down protocol is optimized for primary adipocyte culture. An RNA fragment from androgen receptor's (AR) 3' untranslated region (3'UTR) containing an adenylate-uridylate-rich elementwas used as an example to demonstrate how to retrieve its RBP partner, HuR protein, from adipocyte lystate. The method described here can be applied to detect the interactions between RBPs and noncoding RNAs, as well as between RBPs and coding RNAs.
ISSN: 1940-087X
DOI: 10.3791/54207
Schools: School of Chemical and Biomedical Engineering 
Rights: © 2016 The Author(s) (Journal of Visualized Experiments). This paper was published in Journal of Visualized Experiments and is made available as an electronic reprint (preprint) with permission of The Author(s) (Journal of Visualized Experiments). The published version is available at: []. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SCBE Journal Articles

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