Methods for the Study of Long Noncoding RNA in Cancer Cell Signaling
With the advances in sequencing technology and transcriptome analysis, it is estimated that up to 75 % of the human genome is transcribed into RNAs. This finding prompted intensive investigations on the biological functions of noncoding RNAs and led to very exciting discoveries of microRNAs as important players in disease pathogenesis and therapeutic applications. Research on long noncoding RNAs (lncRNAs) is in its infancy, yet a broad spectrum of biological regulations has been attributed to lncRNAs. Here, we provide a collection of detailed experimental protocols for lncRNA studies, including lncRNA immunoprecipitation, lncRNA pull-down, lncRNA northern blot analysis, lncRNA in situ hybridization, and lncRNA knockdown. We hope that the information included in this chapter can speed up research on lncRNAs biology and eventually lead to the development of clinical applications with lncRNA as novel prognostic markers and therapeutic targets.
Key wordsLong noncoding RNA RNA immunoprecipitation RNA pull-down In situ hybridization Northern blot Short hairpin RNA
This work was supported, in whole or in part, by National Institutes of Health Grant R01CA142776 (L. Zhang), Ovarian Cancer SPORE P50-CA83638-7951 Project 3 (L. Zhang), Department of Defense Grant W81XWH-10-1-0082 (L. Zhang), the Ovarian Cancer Research Fund Tilberis Scholar Award the Basser Research Center grant for BRCA (L. Zhang), and Marsha Rivkin Center for Ovarian Cancer Research (L. Zhang). D. Zhang was supported by the China Scholarship Council.
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