Functional Genomic Strategies for Elucidating Human-Virus Interactions: Will CRISPR Knockout RNAi and Haploid Cells?
Functional Genomic Strategies for Elucidating Human-Virus Interactions: Will CRISPR Knockout RNAi and Haploid Cells?
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DOI：
10.1016/bs.aivir.2015.11.001
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发表时间：
2016
影响因子：
--
通讯作者：
Brass AL
中科院分区：
文献类型：
--
作者：
Perreira JM;Meraner P;Brass AL
在过去的几年里，利用功能丧失功能基因组学产生了大量变革性的人类与病毒相互作用的发现。这些见解极大地扩展了我们对人类致病病毒如何利用我们的细胞进行复制的理解。两项技术一直处于这场基因革命的前沿，即RNA干扰(RNAi)和使用单倍体细胞系的随机逆转录病毒插入突变(单倍体细胞筛选)，前者在很大程度上占主导地位。现在，CRISPR/Cas9系统的尖端基因编辑也已被用于大规模功能基因组学，并有可能取代这些早期的方法。在这里，我们比较和对比了这三种用于阐明宿主-病毒相互作用的筛选方法，概述了它们的主要优点和缺点，包括阵列多个同源RNAi试剂筛选与汇集CRISPR/Cas9人鼻病毒14-人类细胞相互作用筛选的比较，并叙述了每种方法带来的一些值得注意的见解。最后，我们对快速发展的人类病毒功能基因组学领域的前景进行了简要的展望。
Over the last several years a wealth of transformative human–virus interaction discoveries have been produced using loss-of-function functional genomics. These insights have greatly expanded our understanding of how human pathogenic viruses exploit our cells to replicate. Two technologies have been at the forefront of this genetic revolution, RNA interference (RNAi) and random retroviral insertional mutagenesis using haploid cell lines (haploid cell screening), with the former technology largely predominating. Now the cutting edge gene editing of the CRISPR/Cas9 system has also been harnessed for large-scale functional genomics and is poised to possibly displace these earlier methods. Here we compare and contrast these three screening approaches for elucidating host–virus interactions, outline their key strengths and weaknesses including a comparison of an arrayed multiple orthologous RNAi reagent screen to a pooled CRISPR/Cas9 human rhinovirus 14–human cell interaction screen, and recount some notable insights made possible by each. We conclude with a brief perspective on what might lie ahead for the fast evolving field of human–virus functional genomics.
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