Larter Kristina, Yi Bin, Xi Yaguang
Department of Genetics and Stanley S. Scott Cancer Center, Louisiana State University Health Sciences Center, New Orleans, LA, USA.
Methods Mol Biol. 2021;2372:169-177. doi: 10.1007/978-1-0716-1697-0_15.
Long noncoding RNAs (lncRNAs) are a class of RNA transcripts greater than 200 nucleotides in length and makeup a considerable part of the human genome. LncRNAs are well established as crucial players in a myriad of physiological and pathological processes; however, despite their abundance and versatility, the functional characteristics of lncRNAs remain largely unknown predominantly due to the lack of suitable genetic editing strategies. The complexity of their genetic structure and regulation combined with their unique functionality poses several limitations in the application of classic genetic manipulation methods in lncRNA functional studies. Several reports have demonstrated the successful implementation of CRISPR/Cas9 technology in screening and identifying the function of specific lncRNAs. Here, we describe a detailed protocol utilizing CRISPR/Cas9 genetic editing technology for knocking down lncRNAs in vitro.
长链非编码RNA（lncRNAs）是一类长度超过200个核苷酸的RNA转录本，构成了人类基因组的相当一部分。lncRNAs已被确认为众多生理和病理过程中的关键参与者；然而，尽管它们数量众多且功能多样，但lncRNAs的功能特性在很大程度上仍不清楚，主要原因是缺乏合适的基因编辑策略。它们的遗传结构和调控的复杂性，加上其独特的功能，给经典基因操作方法在lncRNA功能研究中的应用带来了一些限制。一些报告已经证明了CRISPR/Cas9技术在筛选和鉴定特定lncRNAs功能方面的成功应用。在这里，我们描述了一种利用CRISPR/Cas9基因编辑技术在体外敲低lncRNAs的详细方案。
