Simon Matthew D
Dept. of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT 06516, USA; Chemical Biology Institute, Yale West Campus, West Haven, CT, 06511, USA.
Biochim Biophys Acta. 2016 Jan;1859(1):121-7. doi: 10.1016/j.bbagrm.2015.09.004. Epub 2015 Sep 14.
Despite mounting evidence of the importance of large non-coding RNAs (lncRNAs) in biological regulation, we still know little about how these lncRNAs function. One approach to understand the function of lncRNAs is to biochemically purify endogenous lncRNAs from fixed cells using complementary oligonucleotides. These hybridization capture approaches can reveal the genomic localization of lncRNAs, as well as the proteins and RNAs with which they interact. To help researchers understand how these tools can uncover lncRNA function, this review discusses the considerations and influences of different parameters, (e.g., crosslinking reagents, oligonucleotide chemistry and hybridization conditions) and controls to avoid artifacts. By examining the application of these tools, this review will highlight the progress and pitfalls of studying lncRNAs using hybridization capture approaches.This article is part of a Special Issue entitled: Clues to long noncoding RNA taxonomy1, edited by Dr. Tetsuro Hirose and Dr. Shinichi Nakagawa.
尽管越来越多的证据表明长链非编码RNA(lncRNA)在生物调控中具有重要作用,但我们对这些lncRNA的功能仍知之甚少。了解lncRNA功能的一种方法是使用互补寡核苷酸从固定细胞中生物化学纯化内源性lncRNA。这些杂交捕获方法可以揭示lncRNA的基因组定位,以及它们相互作用的蛋白质和RNA。为了帮助研究人员了解这些工具如何揭示lncRNA功能,本综述讨论了不同参数(例如交联试剂、寡核苷酸化学和杂交条件)的考虑因素和影响,以及避免假象的对照。通过研究这些工具的应用,本综述将突出使用杂交捕获方法研究lncRNA的进展和陷阱。本文是名为《长链非编码RNA分类线索1》特刊的一部分,由广濑哲郎博士和中川信一博士编辑。