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

Retrovirology

Chen Liang²  Mark A Wainberg²  Fei Guo¹  Jian Li¹  Yann Le Duff²  Rongyue Lei¹  Weijun Zhu¹ 

[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;Departments of Medicine, Microbiology & Immunology, McGill University, Montreal H3A 2B4, Canada

关键词: Genome editing;    CRISPR/Cas9;    Reservoir;    Provirus;    HIV-1;   

Others  :  1162844
DOI  :  10.1186/s12977-015-0150-z

 received in 2014-11-25, accepted in 2015-02-09,  发布年份 2015

【 摘 要 】

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.

【 授权许可】

   
2015 Zhu et al.; licensee BioMed Central.

【 图 表 】

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