Harvard Program in Virology, Harvard Medical School, Boston, MA 02115, USA.
Division of Infectious Disease, Department of Medicine, Brigham & Women's Hospital, Boston, MA 02115, USA.
Viruses. 2018 Jan 30;10(2):55. doi: 10.3390/v10020055.
Clustered regularly interspaced short palindromic repeats (CRISPR) has greatly expanded the ability to genetically probe virus-host interactions. CRISPR systems enable focused or systematic, genomewide studies of nearly all aspects of a virus lifecycle. Combined with its relative ease of use and high reproducibility, CRISPR is becoming an essential tool in studies of the host factors important for viral pathogenesis. Here, we review the use of CRISPR-Cas9 for the loss-of-function analysis of host dependency factors. We focus on the use of CRISPR-pooled screens for the systematic identification of host dependency factors, particularly in Epstein-Barr virus-transformed B cells. We also discuss the use of CRISPR interference (CRISPRi) and gain-of-function CRISPR activation (CRISPRa) approaches to probe virus-host interactions. Finally, we comment on the future directions enabled by combinatorial CRISPR screens.
成簇规律间隔短回文重复序列 (CRISPR) 极大地扩展了遗传探究病毒-宿主相互作用的能力。CRISPR 系统能够集中或系统地、全面地研究病毒生命周期的几乎所有方面。结合其相对易用性和高重复性，CRISPR 正在成为研究病毒发病机制中重要宿主因子的重要工具。在这里，我们回顾了使用 CRISPR-Cas9 进行宿主依赖因子的功能丧失分析。我们专注于使用 CRISPR 池筛选系统地鉴定宿主依赖因子，特别是在 Epstein-Barr 病毒转化的 B 细胞中。我们还讨论了使用 CRISPR 干扰 (CRISPRi) 和功能获得性 CRISPR 激活 (CRISPRa) 方法来探究病毒-宿主相互作用。最后，我们评论了组合 CRISPR 筛选所带来的未来方向。
