Visualization of HIV-1 RNA Transcription from Integrated HIV-1 DNA in Reactivated Latently Infected Cells.
Obiaara B UkahMaritza Puray-ChavezPhilip R TedburyAlon HerschhornJoseph G SodroskiStefan G SarafianosPublished in: Viruses (2018)
We have recently developed the first microscopy-based strategy that enables simultaneous multiplex detection of viral RNA (vRNA), viral DNA (vDNA), and viral protein. Here, we used this approach to study the kinetics of latency reactivation in cells infected with the human immunodeficiency virus (HIV). We showed the transcription of nascent vRNA from individual latently integrated and reactivated vDNA sites appearing earlier than viral protein. We further demonstrated that this method can be used to quantitatively assess the efficacy of a variety of latency reactivating agents. Finally, this microscopy-based strategy was augmented with a flow-cytometry-based approach, enabling the detection of transcriptional reactivation of large numbers of latently infected cells. Hence, these approaches are shown to be suitable for qualitative and quantitative studies of HIV-1 latency and reactivation.
Keyphrases
- human immunodeficiency virus
- antiretroviral therapy
- hiv infected
- hepatitis c virus
- hiv positive
- induced apoptosis
- hiv testing
- hiv aids
- sars cov
- single molecule
- cell cycle arrest
- men who have sex with men
- high resolution
- transcription factor
- high throughput
- label free
- circulating tumor
- cell death
- systematic review
- optical coherence tomography
- real time pcr
- gene expression
- binding protein
- mass spectrometry
- amino acid
- nucleic acid