CRISPRi-based genome-scale identification of functional long noncoding RNA loci in human cells.
CRISPRi-based genome-scale identification of functional long noncoding RNA loci in human cells.
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DOI：
10.1126/science.aah7111
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发表时间：
2017-01-06
期刊：
影响因子：
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通讯作者：
Lim DA
中科院分区：
文献类型：
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作者：
Liu SJ;Horlbeck MA;Cho SW;Birk HS;Malatesta M;He D;Attenello FJ;Villalta JE;Cho MY;Chen Y;Mandegar MA;Olvera MP;Gilbert LA;Conklin BR;Chang HY;Weissman JS;Lim DA
人类基因组产生数千个长的非编码RNA（lncRNA）-长度>200个核苷酸的转录物，其不编码蛋白质。虽然已经揭示了lncRNA的一个子集在正常生物学和疾病中的关键作用，但绝大多数lncRNA的功能仍然未经测试。在这里，我们开发了一种CRISPR干扰（CRISPRi）平台，靶向7种不同细胞系（包括6种转化细胞系和人类诱导多能干细胞（iPSC））中的16，401个lncRNA基因座。大规模筛选鉴定了稳健细胞生长所需的499个lncRNA基因座，其中89%仅在一种细胞类型中显示出生长修饰功能。我们进一步发现，lncRNA敲低可以以细胞类型特异性方式扰乱复杂的转录网络。这些数据强调了许多lncRNA的功能重要性和细胞类型特异性。
The human genome produces thousands of long non-coding RNAs (lncRNAs) – transcripts >200 nucleotides long that do not encode proteins. While critical roles in normal biology and disease have been revealed for a subset of lncRNAs, the function of the vast majority remains untested. Here, we developed a CRISPR interference (CRISPRi) platform targeting 16,401 lncRNA loci in 7 diverse cell lines including 6 transformed cell lines and human induced pluripotent stem cells (iPSCs). Large-scale screening identified 499 lncRNA loci required for robust cellular growth, of which 89% showed growth modifying function exclusively in one cell type. We further found that lncRNA knockdown can perturb complex transcriptional networks in a cell type-specific manner. These data underscore the functional importance and cell type-specificity of many lncRNAs.