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.

随着测序技术和转录组分析的进步，据估计多达75%的人类基因组被转录为RNA。这一发现促使人们对非编码RNA的生物学功能进行深入研究，并在微小RNA作为疾病发病机制和治疗应用中的重要参与者方面有了非常令人兴奋的发现。长链非编码RNA（lncRNAs）的研究尚处于起步阶段，但lncRNAs已被认为具有广泛的生物学调控作用。在此，我们提供了一套用于lncRNA研究的详细实验方案，包括lncRNA免疫沉淀、lncRNA下拉实验、lncRNA northern杂交分析、lncRNA原位杂交以及lncRNA敲低。我们希望本章所包含的信息能够加速lncRNAs生物学的研究，并最终推动以lncRNA作为新型预后标志物和治疗靶点的临床应用的发展。