CRISPR 干扰有效地沉默了卡波西肉瘤相关疱疹病毒感染细胞中的潜伏和裂解病毒基因。
CRISPR Interference Efficiently Silences Latent and Lytic Viral Genes in Kaposi's Sarcoma-Associated Herpesvirus-Infected Cells.
机构信息
Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA 93106, USA.
Neuroscience Research Institute, University of California, Santa Barbara, CA 93106, USA.
出版信息
Viruses. 2021 Apr 28;13(5):783. doi: 10.3390/v13050783.
Uncovering viral gene functions requires the modulation of gene expression through overexpression or loss-of-function. CRISPR interference (CRISPRi), a modification of the CRISPR-Cas9 gene editing technology, allows specific and efficient transcriptional silencing without genetic ablation. CRISPRi has been used to silence eukaryotic and prokaryotic genes at the single-gene and genome-wide levels. Here, we report the use of CRISPRi to silence latent and lytic viral genes, with an efficiency of ~80-90%, in epithelial and B-cells carrying multiple copies of the Kaposi's sarcoma-associated herpesvirus (KSHV) genome. Our results validate CRISPRi for the analysis of KSHV viral elements, providing a functional genomics tool for studying virus-host interactions.
揭示病毒基因功能需要通过过表达或基因功能丧失来调节基因表达。CRISPR 干扰（CRISPRi）是对 CRISPR-Cas9 基因编辑技术的修改，它允许在不进行遗传消融的情况下特异性和有效地进行转录沉默。CRISPRi 已被用于在单细胞和全基因组水平上沉默真核和原核基因。在这里，我们报告了使用 CRISPRi 来沉默潜伏和裂解病毒基因，效率约为 80-90%，在携带卡波济氏肉瘤相关疱疹病毒（KSHV）基因组的多个拷贝的上皮细胞和 B 细胞中。我们的结果验证了 CRISPRi 用于分析 KSHV 病毒元件，为研究病毒-宿主相互作用提供了一种功能基因组学工具。
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