Wang Gang, Zhao Na, Berkhout Ben, Das Atze T
Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Mol Ther. 2016 Mar;24(3):522-6. doi: 10.1038/mt.2016.24. Epub 2016 Jan 22.
Several recent studies demonstrated that the clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease Cas9 can be used for guide RNA (gRNA)-directed, sequence-specific cleavage of HIV proviral DNA in infected cells. We here demonstrate profound inhibition of HIV-1 replication by harnessing T cells with Cas9 and antiviral gRNAs. However, the virus rapidly and consistently escaped from this inhibition. Sequencing of the HIV-1 escape variants revealed nucleotide insertions, deletions, and substitutions around the Cas9/gRNA cleavage site that are typical for DNA repair by the nonhomologous end-joining pathway. We thus demonstrate the potency of CRISPR-Cas9 as an antiviral approach, but any therapeutic strategy should consider the viral escape implications.
最近的几项研究表明,成簇规律间隔短回文重复序列(CRISPR)相关的核酸内切酶Cas9可用于在受感染细胞中对HIV前病毒DNA进行导向RNA(gRNA)介导的序列特异性切割。我们在此证明,通过利用携带Cas9和抗病毒gRNA的T细胞可对HIV-1复制产生显著抑制作用。然而,该病毒能迅速且持续地从这种抑制中逃逸。对HIV-1逃逸变异体进行测序发现,在Cas9/gRNA切割位点周围存在核苷酸插入、缺失和替换,这些是通过非同源末端连接途径进行DNA修复的典型特征。因此,我们证明了CRISPR-Cas9作为一种抗病毒方法的效力,但任何治疗策略都应考虑病毒逃逸的影响。