High throughput generation and characterization of replication-competent clade C transmitter-founder simian human immunodeficiency viruses

Debashis Dutta, Samuel Johnson, Alisha Dalal, Martin J. Deymier, Eric Hunter, Siddappa Nagadenahalli Byrareddy

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Traditional restriction endonuclease-based cloning has been routinely used to generate replication-competent simian-human immunodeficiency viruses (SHIV) and simian tropic HIV (stHIV). This approach requires the existence of suitable restriction sites or the introduction of nucleotide changes to create them. Here, using an In-Fusion cloning technique that involves homologous recombination, we generated SHIVs and stHIVs based on epidemio-logically linked clade C transmitted/founder HIV molecular clones from Zambia. Replacing vif from these HIV molecular clones with vif of SIVmac239 resulted in chimeric genomes used to generate infectious stHIV viruses. Likewise, exchanging HIV env genes and introducing N375 mutations to enhance macaque CD4 binding site and cloned into a SHIVAD8-EO backbone. The generated SHIVs and stHIV were infectious in TZMbl and ZB5 cells, as well as macaque PBMCs. Therefore, this method can replace traditional methods and be a valuable tool for the rapid generation and testing of molecular clones of stHIV and SHIV based on primary clinical isolates will be valuable to generate rapid novel challenge viruses for HIV vaccine/cure studies.

Original languageEnglish (US)
Article numbere0196942
JournalPLoS One
Volume13
Issue number5
DOIs
StatePublished - May 1 2018

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Tropics
Simian Immunodeficiency Virus
Human immunodeficiency virus
Viruses
Transmitters
tropics
Throughput
HIV
Cloning
clones
Macaca
molecular cloning
Genes
AIDS Vaccines
viruses
Zambia
homologous recombination
DNA Restriction Enzymes
restriction endonucleases
mononuclear leukocytes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

High throughput generation and characterization of replication-competent clade C transmitter-founder simian human immunodeficiency viruses. / Dutta, Debashis; Johnson, Samuel; Dalal, Alisha; Deymier, Martin J.; Hunter, Eric; Byrareddy, Siddappa Nagadenahalli.

In: PLoS One, Vol. 13, No. 5, e0196942, 01.05.2018.

Research output: Contribution to journalArticle

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