The CRISPR/Cas9 system inactivates latent HIV-1 proviral DNA

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• 作者：Weijun Zhu (1)
  Rongyue Lei (1)
  Yann Le Duff (2) (3)
  Jian Li (1)
  Fei Guo (1)
  Mark A Wainberg (2) (3)
  Chen Liang (2) (3)
  
  1. MOH Key Laboratory of Systems Biology of Pathogens and AIDS Research Center ; Institute of Pathogen Biology ; Chinese Academy of Medical Sciences & Peking Union Medical College ; Beijing ; 100730 ; PR China
  2. McGill University AIDS Centre ; Lady Davis Institute ; Jewish General Hospital ; Montreal ; H3T 1E2 ; Canada
  3. Departments of Medicine ; Microbiology & Immunology ; McGill University ; Montreal ; H3A 2B4 ; Canada
  
• 关键词：HIV ; 1 ; Provirus ; Reservoir ; CRISPR/Cas9 ; Genome editing
• 刊名：Retrovirology
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：12
• 期：1
• 全文大小：2,366 KB
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• 刊物主题：Virology; Infectious Diseases; Cancer Research;
• 出版者：BioMed Central
• ISSN：1742-4690

文摘

Background 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. Findings 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. Conclusions 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.