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. 2024:2807:31-43.
doi: 10.1007/978-1-0716-3862-0_3.

RNA-FISH for HIV Transcription/Localization Analysis

Pavitra Ramdas  1 , Ajit Chande  2
Affiliations

RNA-FISH for HIV Transcription/Localization Analysis

Pavitra Ramdas et al. Methods Mol Biol. 2024.

Abstract

RNA fluorescence in situ hybridization (FISH) serves as a method for visualizing specific RNA molecules within cells. Its primary utility lies in the observation of messenger RNA (mRNA) molecules associated with particular genes of significance. This technique can also be applied to examine viral transcription and the localization of said transcripts within infected cells. In this context, we provide a comprehensive protocol for the detection, localization, and quantification of HIV-1 transcripts in mammalian cell lines. This encompasses the preparation of required reagents, cellular treatments, visualization, and the subsequent analysis of the data acquired. These parameters play a pivotal role in enhancing our comprehension of the molecular processes during infection, particularly at the crucial transcription phase of the viral life cycle.

Keywords: HIV-1 transcription; Quantification of viral transcripts; RNA-FISH; Transcript localization.

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