التفاصيل البيبلوغرافية
| العنوان: |
High-sensitivity in situ capture of endogenous RNA-protein interactions in fixed cells and primary tissues. |
| المؤلفون: |
Liang, Qishan, Yu, Tao, Kofman, Eric, Jagannatha, Pratibha, Rhine, Kevin, Yee, Brian A., Corbett, Kevin D., Yeo, Gene W. |
| المصدر: |
Nature Communications; 8/16/2024, Vol. 15 Issue 1, p1-16, 16p |
| مصطلحات موضوعية: |
RNA-protein interactions, RNA metabolism, RNA-binding proteins, RNA, WORKFLOW |
| مستخلص: |
RNA-binding proteins (RBPs) have pivotal functions in RNA metabolism, but current methods are limited in retrieving RBP-RNA interactions within endogenous biological contexts. Here, we develop INSCRIBE (IN situ Sensitive Capture of RNA-protein Interactions in Biological Environments), circumventing the challenges through in situ RNA labeling by precisely directing a purified APOBEC1-nanobody fusion to the RBP of interest. This method enables highly specific RNA-binding site identification across a diverse range of fixed biological samples such as HEK293T cells and mouse brain tissue and accurately identifies the canonical binding motifs of RBFOX2 (UGCAUG) and TDP-43 (UGUGUG) in native cellular environments. Applicable to any RBP with available primary antibodies, INSCRIBE enables sensitive capture of RBP-RNA interactions from ultra-low input equivalent to ~5 cells. The robust, versatile, and sensitive INSCRIBE workflow is particularly beneficial for precious tissues such as clinical samples, empowering the exploration of genuine RBP-RNA interactions in RNA-related disease contexts. Retrieving interactions between RNA-binding proteins (RBPs) and RNA in endogenous biological contexts remains challenging. Here, the authors develop INSCRIBE (IN situ Sensitive Capture of RNA-protein Interactions in Biological Environments), which enables highly specific RNA-binding site identification across diverse biological samples. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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