Remsburg Carolyn, Konrad Kalin, Sampilo Nina Faye, Song Jia L
Department of Biological Sciences, University of Delaware, Newark, DE, United States.
Department of Biological Sciences, University of Delaware, Newark, DE, United States.
Methods Cell Biol. 2019;151:323-334. doi: 10.1016/bs.mcb.2018.10.005. Epub 2018 Dec 11.
MicroRNAs (miRNAs) are small non-coding RNAs that repress the translation and reduce the stability of target mRNAs in animal cells. Post-transcriptional regulation mediated by miRNAs is a highly conserved mechanism utilized by organisms throughout phylogeny to fine tune gene expression. We document the approaches used to study the function of a single miRNA and miRNA regulation of biological pathways in the sea urchin embryo. The protocols that are described include selection of miRNA inhibitors, test of miRNA direct targets, and the use of target protector morpholinos to evaluate the impact of miRNA inhibition on its targets. Using the described techniques and strategies, the sea urchin researcher will be able to validate a miRNA's direct targets and evaluate how inhibition of the miRNA affects developmental processes. These results will contribute to our understanding of the regulatory roles of miRNAs in development.
微小RNA（miRNA）是一类小的非编码RNA，可在动物细胞中抑制靶mRNA的翻译并降低其稳定性。由miRNA介导的转录后调控是一种高度保守的机制，生物体在整个系统发育过程中都利用该机制来微调基因表达。我们记录了用于研究单个miRNA功能以及海胆胚胎中生物途径的miRNA调控的方法。所描述的方案包括miRNA抑制剂的选择、miRNA直接靶标的测试，以及使用靶标保护吗啉代寡核苷酸来评估miRNA抑制对其靶标的影响。使用所描述的技术和策略，海胆研究人员将能够验证miRNA的直接靶标，并评估miRNA抑制如何影响发育过程。这些结果将有助于我们理解miRNA在发育中的调控作用。
