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

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

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

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 (rAAV9) 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 rAAV9: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 rAAV9 vector in a range of cells and tissues that harbor integrated copies of viral DNA.

Original languageEnglish (US)
Pages (from-to)690-695
Number of pages6
JournalGene Therapy
Volume23
Issue number8-9
DOIs
StatePublished - Aug 1 2016

Fingerprint

Guide RNA
Viral DNA
HIV-1
Dependovirus
Clustered Regularly Interspaced Short Palindromic Repeats
DNA
gag Genes
Transgenic Rats
Transgenic Mice
Genome
Lymphocytes
Viral Genes
Endonucleases
Tail
Veins
Spleen
Gene Editing
T-Lymphocytes
Kidney
Gene Expression

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Kaminski, R., Bella, R., Yin, C., Otte, J., Ferrante, P., Gendelman, H. E., ... Khalili, K. (2016). Excision of HIV-1 DNA by gene editing: A proof-of-concept in vivo study. Gene Therapy, 23(8-9), 690-695. https://doi.org/10.1038/gt.2016.41

Excision of HIV-1 DNA by gene editing : A proof-of-concept in vivo study. / Kaminski, R.; Bella, R.; Yin, C.; Otte, J.; Ferrante, P.; Gendelman, Howard Eliot; Li, H.; Booze, R.; Gordon, J.; Hu, W.; Khalili, K.

In: Gene Therapy, Vol. 23, No. 8-9, 01.08.2016, p. 690-695.

Research output: Contribution to journalArticle

Kaminski, R, Bella, R, Yin, C, Otte, J, Ferrante, P, Gendelman, HE, Li, H, Booze, R, Gordon, J, Hu, W & Khalili, K 2016, 'Excision of HIV-1 DNA by gene editing: A proof-of-concept in vivo study', Gene Therapy, vol. 23, no. 8-9, pp. 690-695. https://doi.org/10.1038/gt.2016.41
Kaminski, R. ; Bella, R. ; Yin, C. ; Otte, J. ; Ferrante, P. ; Gendelman, Howard Eliot ; Li, H. ; Booze, R. ; Gordon, J. ; Hu, W. ; Khalili, K. / Excision of HIV-1 DNA by gene editing : A proof-of-concept in vivo study. In: Gene Therapy. 2016 ; Vol. 23, No. 8-9. pp. 690-695.
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