IFI16 knockdown in primary HIV-1 target cells

STAR Protoc. 2020 Dec 19;2(1):100236. doi: 10.1016/j.xpro.2020.100236. eCollection 2021 Mar 19.

Abstract

IFI16 is an important player of the host intrinsic immune response. Among others, it has been reported to sense intermediate products of HIV-1 reverse transcription in the cytosol and to sequester the transcription factor Sp1 in the nucleus to attenuate viral gene expression. Here, we present three different methods to reduce IFI16 protein expression levels in HIV-1 primary target cells. These techniques can be adapted for the investigation of other cellular factors in primary macrophages and CD4+ T lymphocytes. For complete details on the use and execution of this protocol, please refer to Hotter et al. (2019) and Bosso et al. (2020).

Keywords: Immunology; Molecular Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD4-Positive T-Lymphocytes* / metabolism
  • CD4-Positive T-Lymphocytes* / virology
  • Gene Knockdown Techniques*
  • HIV Infections*
  • HIV-1 / metabolism*
  • Humans
  • Macrophages* / metabolism
  • Macrophages* / virology
  • Nuclear Proteins* / genetics
  • Nuclear Proteins* / metabolism
  • Phosphoproteins* / genetics
  • Phosphoproteins* / metabolism

Substances

  • Nuclear Proteins
  • Phosphoproteins
  • IFI16 protein, human