Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, NY, United States of America.
Department of Molecular and Cellular Physiology, Albany Medical College, Albany, NY, United States of America.
Vascul Pharmacol. 2019 Mar;114:1-12. doi: 10.1016/j.vph.2019.02.004. Epub 2019 Feb 27.
Next generation sequencing has uncovered a trove of short noncoding RNAs (e.g., microRNAs) and long noncoding RNAs (lncRNAs) that act as molecular rheostats in the control of diverse homeostatic processes. Meanwhile, the tsunamic emergence of clustered regularly interspaced short palindromic repeats (CRISPR) editing has transformed our influence over all DNA-carrying entities, heralding global CRISPRization. This is evident in biomedical research where the ease and low-cost of CRISPR editing has made it the preferred method of manipulating the mouse genome, facilitating rapid discovery of genome function in an in vivo context. Here, CRISPR genome editing components are updated for elucidating lncRNA function in mice. Various strategies are highlighted for understanding the function of lncRNAs residing in intergenic sequence space, as host genes that harbor microRNAs or other genes, and as natural antisense, overlapping or intronic genes. Also discussed is CRISPR editing of mice carrying human lncRNAs as well as the editing of competing endogenous RNAs. The information described herein should assist labs in the rigorous design of experiments that interrogate lncRNA function in mice where complex disease processes can be modeled thus accelerating translational discovery.
下一代测序技术揭示了大量短非编码 RNA(例如 microRNAs)和长非编码 RNA(lncRNAs),它们作为分子变阻器,控制着多种体内平衡过程。与此同时,成簇规律间隔短回文重复序列(CRISPR)编辑的海啸式出现改变了我们对所有携带 DNA 的实体的影响,预示着全球 CRISPR 化。这在生物医学研究中显而易见,CRISPR 编辑的简便性和低成本使其成为操纵小鼠基因组的首选方法,促进了在体内环境中快速发现基因组功能。在这里,CRISPR 基因组编辑组件被更新,以阐明小鼠中的 lncRNA 功能。强调了各种策略,用于了解位于基因间序列空间、作为 miRNA 或其他基因的宿主基因、以及作为天然反义、重叠或内含子基因的 lncRNA 的功能。还讨论了携带人类 lncRNAs 的小鼠的 CRISPR 编辑以及竞争内源性 RNA 的编辑。本文所述的信息应有助于实验室在严格设计实验中,探究小鼠中的 lncRNA 功能,从而可以模拟复杂的疾病过程,加速转化发现。