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Construction and Characterization of Infectious Molecular Clones of HIV-1 CRF63_02A6

Daria P Zyryanova1, Alexei V Totmenin1, Natalia V Bogacheva1

  • 1Department of Retroviruses, State Research Center of Virology and Biotechnology "Vector", Koltsovo, Russia.

AIDS Research and Human Retroviruses
|September 5, 2019
PubMed

Insights

Researchers constructed infectious molecular clones of the prevalent HIV-1 CRF63_02A6 variant circulating in Siberia. These well-characterized clones facilitate further study of this significant HIV subtype.

Area of Science:

  • Virology
  • Genetics
  • Molecular Biology

Background:

  • The HIV-1 CRF63_02A6 subtype is the predominant genetic variant among HIV-infected individuals in the Siberian Federal District, Russia.
  • Understanding the characteristics of prevalent HIV strains is crucial for developing effective treatment and prevention strategies.

Purpose of the Study:

  • To construct and characterize infectious molecular clones (IMCs) of the HIV-1 CRF63_02A6 subtype.
  • To provide tools for basic and applied research on this prevalent Russian HIV-1 variant.

Main Methods:

  • Construction of near full-length proviral HIV-1 sequences (pT11.17 and pMtBs.18) from 2015 and 2017 isolates.
  • Production of IMC virions via HEK293T cell transfection and assessment of viral growth in peripheral blood mononuclear cell cultures.
  • Determination of viral tropism using genotypic (Geno2pheno) and phenotypic assays in MT-2, U87-CD4-CCR5, and U87-CD4-CXCR4 cell lines.

Main Results:

  • Stable replication of CRF63_02A6 IMCs was achieved, with significant increases in p24 concentration and viral protein accumulation.
  • Genotypic and phenotypic analyses confirmed the R5-tropic and X4-tropic nature of the constructed IMCs.
  • The generated IMCs represent viable tools for studying HIV-1 CRF63_02A6.

Conclusions:

  • Successfully constructed and characterized R5-tropic and X4-tropic HIV-1 CRF63_02A6 infectious molecular clones.
  • These IMCs are stable and replicate efficiently in cell culture, validating their utility.
  • The developed IMCs offer valuable resources for advancing research on HIV-1 CRF63_02A6 in Russia and beyond.
Keywords:
CRF63_02A6HIV-1 isolatesinfectious molecular clone

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