Tumor Initiation and Maintenance Program, NCI-designated Cancer Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, 92037, USA.
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA, 92121, USA.
Sci Rep. 2019 Feb 20;9(1):2408. doi: 10.1038/s41598-018-36750-0.
Despite essential roles played by long noncoding RNAs (lncRNAs) in development and disease, methods to determine lncRNA cis-elements are lacking. Here, we developed a screening method named "Tiling CRISPR" to identify lncRNA functional domains. Using this approach, we identified Xist A-Repeats as the silencing domain, an observation in agreement with published work, suggesting Tiling CRISPR feasibility. Mechanistic analysis suggested a novel function for Xist A-repeats in promoting Xist transcription. Overall, our method allows mapping of lncRNA functional domains in an unbiased and potentially high-throughput manner to facilitate the understanding of lncRNA functions.
尽管长非编码 RNA（lncRNAs）在发育和疾病中发挥着重要作用，但确定 lncRNA 顺式元件的方法仍很缺乏。在这里，我们开发了一种名为“嵌合 CRISPR”的筛选方法来鉴定 lncRNA 的功能结构域。使用这种方法，我们鉴定了 Xist A 重复序列作为沉默结构域，这一观察结果与已发表的工作一致，表明嵌合 CRISPR 是可行的。机制分析表明 Xist A 重复序列在促进 Xist 转录方面具有新的功能。总的来说，我们的方法允许以一种无偏和潜在高通量的方式对 lncRNA 功能结构域进行映射，从而促进对 lncRNA 功能的理解。
