CRISPR/Cas9 Inhibits Multiple Steps of HIV-1 Infection
CRISPR/Cas9 Inhibits Multiple Steps of HIV-1 Infection
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CRISPR/Cas9 抑制 HIV-1 感染的多个步骤
DOI：
10.1089/hum.2018.018
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发表时间：
2018
影响因子：
4.2
通讯作者：
Guo Fei
中科院分区：
文献类型：
--
作者：
Yin Lijuan;Hu Siqi;Mei Shan;Sun Hong;Xu Fengwen;Li Jian;Zhu Weijun;Liu Xiaoman;Zhao Fei;Zhang Di;Cen Shan;Liang Chen;Guo Fei
CRISPR/Cas9是一种适应性免疫系统，细菌和古菌通过在DNA中产生位点特异的双链断裂来进化来抵抗入侵的病毒和质粒DNA。本研究通过针对病毒长末端重复序列和基因编码序列，测试了该基因编辑系统对人类免疫缺陷病毒1型(HIV-1)感染的抑制作用。观察到Cas9/gRNA对HIV-1感染的强烈抑制作用，这不仅是由于Cas9裂解导致插入和缺失(Indels)到病毒DNA中，而且是由于晚期病毒DNA产物和整合病毒DNA水平的显著下降。后一种缺陷可能反映了病毒DNA的降解，在Cas9切割后没有立即修复。进一步观察到，当Cas9单独定位于细胞质时，对HIV-1的抑制作用与核Cas9一样强，只是细胞质Cas9不作用于整合的HIV-1DNA，因此不能用于切除潜伏的前病毒。综上所述，这些结果表明Cas9/gRNA能够在细胞质和细胞核中靶向和编辑HIV-1DNA。Cas9对HIV-1的抑制作用既与病毒DNA中的Indels有关，又与其降低病毒DNA水平有关。
CRISPR/Cas9 is an adaptive immune system where bacteria and archaea have evolved to resist the invading viruses and plasmid DNA by creating site-specific double-strand breaks in DNA. This study tested this gene editing system in inhibiting human immunodeficiency virus type 1 (HIV-1) infection by targeting the viral long terminal repeat and the gene coding sequences. Strong inhibition of HIV-1 infection by Cas9/gRNA was observed, which resulted not only from insertions and deletions (indels) that were introduced into viral DNA due to Cas9 cleavage, but also from the marked decrease in the levels of the late viral DNA products and the integrated viral DNA. This latter defect might have reflected the degradation of viral DNA that has not been immediately repaired after Cas9 cleavage. It was further observed that Cas9, when solely located in the cytoplasm, inhibits HIV-1 as strongly as the nuclear Cas9, except that the cytoplasmic Cas9 does not act on the integrated HIV-1 DNA and thus cannot be used to excise the latent provirus. Together, the results suggest that Cas9/gRNA is able to target and edit HIV-1 DNA both in the cytoplasm and in the nucleus. The inhibitory effect of Cas9 on HIV-1 is attributed to both the indels in viral DNA and the reduction in the levels of viral DNA.