Characterization of Long Noncoding RNA-Associated Proteins by RNA-Immunoprecipitation
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. RNA-immunoprecipitation (RNA-IP) is a technique of detecting the association of individual proteins with specific RNA molecules in vivo. It can be used to investigate lncRNA-protein interaction and identify lncRNAs that bind to a protein of interest. Here we describe the protocol of this assay with detailed materials and methods.
Key wordsLong noncoding RNA RNA-binding protein RNA-immunoprecipitation
This work was supported, in whole or in part, by the Basser Research Center for BRCA, the NIH (R01CA142776, R01CA190415, P50CA083638, P50CA174523), the Ovarian Cancer Research Fund (XH), the Breast Cancer Alliance, Foundation for Women’s Cancer (XH), and the Marsha Rivkin Center for Ovarian Cancer Research.