一种整合的全基因组 CRISPRa 方法，用于赋予耐药性中长链非编码 RNA 的功能。
An Integrated Genome-wide CRISPRa Approach to Functionalize lncRNAs in Drug Resistance.
机构信息
Cancer Research Institute, Beth Israel Deaconess Cancer Center, Department of Medicine and Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Ludwig Center at Harvard, Harvard Medical School, Boston, MA, USA.
Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA; Department of Pathology and Cell Biology, Columbia University College of Physicians and Surgeons, New York, NY 10032, USA; Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Columbia University College of Physicians and Surgeons, New York, NY 10032, USA.
出版信息
Cell. 2018 Apr 19;173(3):649-664.e20. doi: 10.1016/j.cell.2018.03.052.
Resistance to chemotherapy plays a significant role in cancer mortality. To identify genetic units affecting sensitivity to cytarabine, the mainstay of treatment for acute myeloid leukemia (AML), we developed a comprehensive and integrated genome-wide platform based on a dual protein-coding and non-coding integrated CRISPRa screening (DICaS). Putative resistance genes were initially identified using pharmacogenetic data from 760 human pan-cancer cell lines. Subsequently, genome scale functional characterization of both coding and long non-coding RNA (lncRNA) genes by CRISPR activation was performed. For lncRNA functional assessment, we developed a CRISPR activation of lncRNA (CaLR) strategy, targeting 14,701 lncRNA genes. Computational and functional analysis identified novel cell-cycle, survival/apoptosis, and cancer signaling genes. Furthermore, transcriptional activation of the GAS6-AS2 lncRNA, identified in our analysis, leads to hyperactivation of the GAS6/TAM pathway, a resistance mechanism in multiple cancers including AML. Thus, DICaS represents a novel and powerful approach to identify integrated coding and non-coding pathways of therapeutic relevance.
化疗耐药在癌症死亡率中起着重要作用。为了确定影响阿糖胞苷敏感性的遗传单元，阿糖胞苷是治疗急性髓细胞白血病（AML）的主要药物，我们开发了一种基于双蛋白编码和非编码整合 CRISPRa 筛选（DICaS）的全面综合基因组平台。最初使用来自 760 个人类泛癌细胞系的药物遗传学数据来鉴定潜在的耐药基因。随后，通过 CRISPR 激活对编码和长非编码 RNA（lncRNA）基因进行了全基因组功能特征分析。为了评估 lncRNA 的功能，我们开发了一种靶向 14701 个 lncRNA 基因的 CRISPR 激活 lncRNA（CaLR）策略。计算和功能分析鉴定了新的细胞周期、存活/凋亡和癌症信号基因。此外，我们的分析中鉴定的 GAS6-AS2 lncRNA 的转录激活导致 GAS6/TAM 途径的过度激活，这是包括 AML 在内的多种癌症的一种耐药机制。因此，DICaS 代表了一种识别治疗相关的整合编码和非编码途径的新的、强大的方法。
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