Institut Curie, PSL Research University, CNRS UMR3215, INSERM U934, Paris, France.
IUBMB Life. 2023 Jun;75(6):458-470. doi: 10.1002/iub.2690. Epub 2022 Dec 7.
LncRNAs represent an abundant group of noncoding transcripts, some of which carry out important regulatory functions. To survey the biological and molecular roles of lncRNAs, reliable strategies for their genetic inactivation are required. Several lncRNA features make them challenging to target by genome editing. First, lncRNA loci often span large genomic distances. As such, full or partial deletion alleles are not always easy to generate and interpret as they might affect DNA regulatory elements. Second, in contrast to proteins, lncRNA transcripts are usually resistant to the minimally invasive approach of point substitutions. Third, lncRNA sequences exhibit rapid evolutionary turnover, impeding prediction and targeting of the specific functional sequence elements. Nonetheless, advances in genome editing and comparative genomics have expanded the repertoire of genetic strategies to dissect lncRNA functions in model organisms and cell lines. In this review, we discuss several approaches that have been used to generate lncRNA mutant alleles, focusing on vertebrate lncRNAs. We also briefly highlight comparative genomics approaches to identify conserved lncRNA sequence motifs, which represent attractive target sequences to abrogate lncRNA functions and to pinpoint functional contributions of these elements.
长链非编码 RNA 是一类丰富的非编码转录本，其中一些具有重要的调节功能。为了研究长链非编码 RNA 的生物学和分子作用，需要可靠的遗传失活策略。几个长链非编码 RNA 的特征使得它们难以通过基因组编辑进行靶向。首先，长链非编码 RNA 基因座通常跨越很大的基因组距离。因此，完全或部分缺失等位基因并不总是容易产生和解释，因为它们可能影响 DNA 调节元件。其次，与蛋白质不同，长链非编码 RNA 转录本通常对最小侵入性的点突变方法具有抗性。第三，长链非编码 RNA 序列表现出快速的进化更替，阻碍了特定功能序列元件的预测和靶向。尽管如此，基因组编辑和比较基因组学的进展扩大了遗传策略的范围，以在模式生物和细胞系中剖析长链非编码 RNA 的功能。在这篇综述中，我们讨论了几种用于产生长链非编码 RNA 突变等位基因的方法，重点是脊椎动物长链非编码 RNA。我们还简要介绍了比较基因组学方法，以识别保守的长链非编码 RNA 序列基序，这些基序是消除长链非编码 RNA 功能和确定这些元件的功能贡献的有吸引力的靶序列。
