Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture
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...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2017
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| Online Access: | https://hdl.handle.net/10356/83049 http://hdl.handle.net/10220/42373 |
| _version_ | 1811694475804672000 |
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| author | Bai, Qianfan Bai, Zhiqiang Xu, Shaohai Sun, Lei |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Bai, Qianfan Bai, Zhiqiang Xu, Shaohai Sun, Lei |
| author_sort | Bai, Qianfan |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T07:08:10Z |
| format | Journal Article |
| id | ntu-10356/83049 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:08:10Z |
| publishDate | 2017 |
| record_format | dspace |
| spelling | ntu-10356/830492023-12-29T06:51:39Z Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture Bai, Qianfan Bai, Zhiqiang Xu, Shaohai Sun, Lei School of Chemical and Biomedical Engineering Molecular Biology RNA-binding protein 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. NMRC (Natl Medical Research Council, S’pore) Published version 2017-05-11T06:13:01Z 2019-12-06T15:10:51Z 2017-05-11T06:13:01Z 2019-12-06T15:10:51Z 2016 Journal Article 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-. 1940-087X https://hdl.handle.net/10356/83049 http://hdl.handle.net/10220/42373 10.3791/54207 en Journal of Visualized Experiments © 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: [http://dx.doi.org/10.3791/54207]. 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. 7 p. application/pdf |
| spellingShingle | Molecular Biology RNA-binding protein Bai, Qianfan Bai, Zhiqiang Xu, Shaohai Sun, Lei Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture |
| title | Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture |
| title_full | Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture |
| title_fullStr | Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture |
| title_full_unstemmed | Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture |
| title_short | Detection of RNA-binding Proteins by RNA Pull-down in Adipocyte Culture |
| title_sort | detection of rna binding proteins by rna pull down in adipocyte culture |
| topic | Molecular Biology RNA-binding protein |
| url | https://hdl.handle.net/10356/83049 http://hdl.handle.net/10220/42373 |
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