Multiplex single-cell visualization of nucleic acids and protein during HIV infection

• DOI: 10.1038/s41467-017-01693-z
• Publication Date: 2017-11-27T10:48:38Z
• Authors: Maritza Puray-Chavez; Philip R. Tedbury; Andrew D. Huber; Obiaara B. Ukah; Vincent Yapo; Dandan Liu; Juan Ji; Jennifer J. Wolf; Alan N. Engelman; Stefan G. Sarafianos
• Keywords: Multiplex; Transcription

Technical limitations in simultaneous microscopic visualization of RNA, DNA, and proteins HIV have curtailed progress this field. To address need we develop a microscopy approach, multiplex immunofluorescent cell-based detection RNA Protein (MICDDRP), which is based on branched DNA situ hybridization technology. MICDDRP enables single-cell (a) spliced unspliced (b) cytoplasmic nuclear (c) Gag. We use to visualize incoming capsid cores containing and/or nascent follow reverse transcription kinetics. also report transcriptional "bursts" from integrated proviral concomitant HIV-1, HIV-2 co-infected cells. can be used simultaneously detect multiple viral nucleic acid intermediates, characterize the effects host factors or drugs steps life cycle, its reactivation latent state, thus facilitating development antivirals latency reactivating agents.