Archive for July 22, 2014

RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES

Wenhui Hua,1,2, Rafal Kaminskia,1, Fan Yanga, Yonggang Zhanga, Laura Cosentinoa, Fang Lia, Biao Luob, David Alvarez-Carbonellc, Yoelvis Garcia-Mesac, Jonathan Karnc, Xianming Mod, and Kamel Khalilia,2

Author Affiliations

aDepartment of Neuroscience, Center for Neurovirology and The Comprehensive NeuroAIDS Center, Temple University School of Medicine, Philadelphia, PA 19140;

bCancer Genome Institute, Fox Chase Cancer Center, Temple University School of Medicine, Philadelphia, PA 19111;

cDepartment of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH 44106; and

dLaboratory of Stem Cell Biology, State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China

Significance

For more than three decades since the discovery of HIV-1, AIDS remains a major public health problem affecting greater than 35.3 million people worldwide. Current antiretroviral therapy has failed to eradicate HIV-1, partly due to the persistence of viral reservoirs. RNA-guided HIV-1 genome cleavage by the Cas9 technology has shown promising efficacy in disrupting the HIV-1 genome in latently infected cells, suppressing viral gene expression and replication, and immunizing uninfected cells against HIV-1 infection. These properties may provide a viable path toward a permanent cure for AIDS, and provide a means to vaccinate against other pathogenic viruses. Given the ease and rapidity of Cas9/guide RNA development, personalized therapies for individual patients with HIV-1 variants can be developed instantly.

Abstract

AIDS remains incurable due to the permanent integration of HIV-1 into the host genome, imparting risk of viral reactivation even after antiretroviral therapy. New strategies are needed to ablate the viral genome from latently infected cells, because current methods are too inefficient and prone to adverse off-target effects. To eliminate the integrated HIV-1 genome, we used the Cas9/guide RNA (gRNA) system, in single and multiplex configurations. We identified highly specific targets within the HIV-1 LTR U3 region that were efficiently edited by Cas9/gRNA, inactivating viral gene expression and replication in latently infected microglial, promonocytic, and T cells. Cas9/gRNAs caused neither genotoxicity nor off-target editing to the host cells, and completely excised a 9,709-bp fragment of integrated proviral DNA that spanned from its 5′ to 3′ LTRs. Furthermore, the presence of multiplex gRNAs within Cas9-expressing cells prevented HIV-1 infection. Our results suggest that Cas9/gRNA can be engineered to provide a specific, efficacious prophylactic and therapeutic approach against AIDS.

PDF

http://www.pnas.org/content/early/2014/07/17/1405186111.full.pdf+html

Advertisements

July 22, 2014 at 2:20 pm


Calendar

July 2014
M T W T F S S
« Jun   Aug »
 123456
78910111213
14151617181920
21222324252627
28293031  

Posts by Month

Posts by Category