Zhu Weijun, Lei Rongyue, Le Duff Yann, Li Jian, Guo Fei, Wainberg Mark A, Liang Chen
Retrovirology. 2015 Feb 27;12:22. doi: 10.1186/s12977-015-0150-z.
Highly active antiretroviral therapy (HAART) has transformed HIV-1 infection from a deadly disease to a manageable chronic illness, albeit does not provide a cure. The recently developed genome editing system called CRISPR/Cas9 offers a new tool to inactivate the integrated latent HIV-1 DNA and may serve as a new avenue toward cure.
We tested 10 sites in HIV-1 DNA that can be targeted by CRISPR/Cas9. The engineered CRISPR/Cas9 system was introduced into the JLat10.6 cells that are latently infected by HIV-1. The sequencing results showed that each target site in HIV-1 DNA was efficiently mutated by CRISPR/Cas9 with the target site in the second exon of Rev (called T10) exhibiting the highest degree of mutation. As a result, HIV-1 gene expression and virus production were significantly diminished with T10 causing a 20-fold reduction.
The CRISPR/Cas9 complex efficiently mutates and deactivates HIV-1 proviral DNA in latently infected Jurkat cells. Our results also revealed a highly efficient Cas9 target site within the second exon of Rev that represents a promising target to be further explored in the CRISPR/Cas9-based cure strategy.
高效抗逆转录病毒疗法（HAART）已将HIV-1感染从一种致命疾病转变为一种可控制的慢性病，尽管无法治愈。最近开发的名为CRISPR/Cas9的基因组编辑系统提供了一种使整合的潜伏HIV-1 DNA失活的新工具，可能成为治愈的新途径。
我们测试了HIV-1 DNA中10个可被CRISPR/Cas9靶向的位点。将工程化的CRISPR/Cas9系统导入被HIV-1潜伏感染的JLat10.6细胞中。测序结果表明，CRISPR/Cas9能有效使HIV-1 DNA中的每个靶位点发生突变，其中Rev第二个外显子中的靶位点（称为T10）突变程度最高。结果，HIV-1基因表达和病毒产生显著减少，T10导致其减少20倍。
CRISPR/Cas9复合物能有效使潜伏感染的Jurkat细胞中的HIV-1前病毒DNA发生突变并失活。我们的结果还揭示了Rev第二个外显子内一个高效的Cas9靶位点，这是在基于CRISPR/Cas9的治愈策略中有待进一步探索的一个有前景的靶点。
