Institute of Pharmacology and Toxicology, Technical University Munich (TUM), Munich, Germany.
Department of Biochemistry and BioFrontiers, University of Colorado Boulder, Boulder, CO, USA.
Nat Rev Genet. 2022 Apr;23(4):229-243. doi: 10.1038/s41576-021-00427-8. Epub 2021 Nov 26.
Genome-wide sequencing has led to the discovery of thousands of long non-coding RNA (lncRNA) loci in the human genome, but evidence of functional significance has remained controversial for many lncRNAs. Genetically engineered model organisms are considered the gold standard for linking genotype to phenotype. Recent advances in CRISPR-Cas genome editing have led to a rapid increase in the use of mouse models to more readily survey lncRNAs for functional significance. Here, we review strategies to investigate the physiological relevance of lncRNA loci by highlighting studies that have used genetic mouse models to reveal key in vivo roles for lncRNAs, from fertility to brain development. We illustrate how an investigative approach, starting with whole-gene deletion followed by transcription termination and/or transgene rescue strategies, can provide definitive evidence for the in vivo function of mammalian lncRNAs.
全基因组测序已经发现了人类基因组中数千个长非编码 RNA（lncRNA）基因座，但许多 lncRNA 的功能意义仍存在争议。遗传工程模型生物被认为是将基因型与表型联系起来的金标准。CRISPR-Cas 基因组编辑的最新进展使得更多地使用小鼠模型来更易于研究 lncRNA 的功能意义成为可能。在这里，我们通过强调使用遗传小鼠模型揭示 lncRNA 在从生育到大脑发育等方面的关键体内作用的研究，来综述调查 lncRNA 基因座生理相关性的策略。我们说明了一种从全基因缺失开始，然后是转录终止和/或转基因拯救策略的研究方法，如何为哺乳动物 lncRNA 的体内功能提供明确的证据。
