利用全基因组CRISPRi/a筛选系统性鉴定lncRNA对人类神经元分化、存活及神经突生长的影响

人类神经系统中存在大量特异性表达的长链非编码RNA（lncRNA）。已有研究表明lncRNA对于人类神经系统的发育和正常生理功能起着至关重要的作用。然而，绝大部分lncRNA在神经系统中的具体功能尚不清楚。虽然已有众多转录组学研究揭示出lncRNA表达水平的信息，但是对于lncRNA在人类神经系统中的功能目前尚无系统性研究。申请人前期工作将CRISPRi/a基因表达干扰系统与人类诱导多能干细胞（iPSC）神经分化技术相结合，开发出了在人类神经元中进行全基因组水平基因功能研究的平台。本课题将利用这一遗传筛选平台，系统性地研究神经元特异的lncRNA对于神经元分化、存活以及神经突生长的影响，初步绘制lncRNA在人类神经元中的功能图谱。对于鉴定出的关键lncRNA，我们将深入阐述其分子细胞生物学机制。本研究将对系统性研究lncRNA在神经系统及其他组织细胞类型中的功能提供新的思路。

A large number of long non-coding RNAs (lncRNAs) are specifically expressed in the human nervous system. Previous studies have revealed that lncRNAs can play pivotal roles in regulating the development and normal physiological function of the nervous system. However, functions of the vast majority lncRNAs are still unknown. Although a number of transcriptome studies have identified expression changes of lncRNAs under different conditions, systematic functional study of lncRNA in the human nervous system is still lacking. We have previously developed a genetic screen platform in human neurons by combining CRISPRi/a-based genetic perturbation tools with human iPSC neuronal differentiation methods. This platform allows us to systematically study gene function in human neurons on a genome-wide scale. In this study, we propose to apply this screening platform to systematically study the effects of neuronal-specific lncRNA on neuronal differentiation, neuronal survival and neurite outgrowth, resulting in a preliminary functional landscape of lncRNAs in human neurons. We will also elucidate the cellular and molecular mechanisms of key lncRNAs identified above. This study will provide a new paradigm for the study of lncRNA function in the nervous system and other tissue/cell types.