Journal Article

Excision of HIV-1 DNA by gene editing: a proof-of-concept in vivo study

Booze, R, Kaminski, R, Yin, C, Li, H, Gordon, J, Hu, W, Khalili, K, Ferrante, P, Bella, R, Otte, J, Gendelman, H E

2016

Overview

A CRISPR/Cas9 gene editing strategy has been remarkable in excising segments of integrated HIV-1 DNA sequences from the genome of latently infected human cell lines and by introducing InDel mutations, suppressing HIV-1 replication in patient-derived CD4+ T-cells, ex vivo . Here, we employed a short version of the Cas9 endonuclease, saCas9, together with a multiplex of guide RNAs (gRNAs) for targeting the viral DNA sequences within the 5′-LTR and the Gag gene for removing critically important segments of the viral DNA in transgenic mice and rats encompassing the HIV-1 genome. Tail-vein injection of transgenic mice with a recombinant Adeno-associated virus 9 (rAAV 9 ) vector expressing saCas9 and the gRNAs, rAAV:saCas9/gRNA, resulted in the cleavage of integrated HIV-1 DNA and excision of a 978 bp DNA fragment spanning between the LTR and Gag gene in the spleen, liver, heart, lung and kidney as well as in the circulating lymphocytes. Retro-orbital inoculation of rAAV 9 :saCas9/gRNA in transgenic rats eliminated a targeted segment of viral DNA and substantially decreased the level of viral gene expression in circulating blood lymphocytes. The results from the proof-of-concept studies, for the first time, demonstrate the in vivo eradication of HIV-1 DNA by CRISPR/Cas9 on delivery by an rAAV 9 vector in a range of cells and tissues that harbor integrated copies of viral DNA.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 45/41 / 64/110 / 64/60 / 692/699/1541 / 692/699/255 / Adeno-associated virus / AIDS treatment / Animals / Biomedical and Life Sciences / Biomedicine / Care and treatment / CD4 antigen / CD4-Positive T-Lymphocytes - metabolism / Cell Biology / Cell lines / Cells, Cultured / CRISPR / CRISPR-Cas Systems / Deoxyribonucleic acid / Dependovirus - genetics / DNA / DNA, Viral - genetics / Endonuclease / Gag protein / Gene Editing - methods / Gene Expression / Gene Products, gag - genetics / Gene Targeting - methods / Gene Therapy / Genetic aspects / Genetic engineering / Genome editing / Genomes / gRNA / Health aspects / HIV / HIV (Viruses) / HIV infection / HIV-1 - genetics / Human Genetics / Human immunodeficiency virus / Human immunodeficiency virus 1 / Innovations / Inoculation / Kidney - metabolism / Kidneys / Latent infection / Lentivirus / Liver - metabolism / Lung - metabolism / Lymphocytes / Lymphocytes T / Methods / Mice / Myocardium - metabolism / Nanotechnology / Nucleotide sequence / Rats / short-communication / Spleen / Transgenic animals / Transgenic mice