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.
Abramson Family Cancer Research Institute, School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA.
Yi
Y
Feng
Xiaowen
X
Hu
Youyou
Y
Zhang
Dongmei
D
Zhang
Chunsheng
C
Li
Lin
L
Zhang
D019921:Biotinylation; D015152:Blotting, Northern; D063646:Carcinogenesis; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics; D055785:Gene Knockdown Techniques; D005821:Genetic Techniques; D006801:Humans; D047468:Immunoprecipitation; D017403:In Situ Hybridization; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology*; D062085:RNA, Long Noncoding / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D014158:Transcription, Genetic
D062085:RNA, Long Noncoding; D034741:RNA, Small Interfering