INVESTIGATION OF HIV-1  ANTISENSE TRANSCRIPT EXPRESSION IN VIVO

Sklutuis, Rachel

INVESTIGATION OF HIV-1 ANTISENSE TRANSCRIPT EXPRESSION IN VIVO

dc.contributor.author	Sklutuis, Rachel
dc.contributor.department	Hood College Department of Biology	en_US
dc.contributor.program	Biomedical Science	en_US
dc.date.accessioned	2022-04-15T13:26:55Z
dc.date.available	2022-04-15T13:26:55Z
dc.description.abstract	Natural antisense transcripts (NATs) are a class of RNA molecules that function in regulating gene expression and are transcribed from the opposite strand of some protein-coding genes. In Human Immunodeficiency virus 1 (HIV-1), in vitro experiments have shown that expression of a NAT called Ast promotes the establishment and maintenance of viral latency through polycomb repressive complex 2 (PRC2)-induced epigenetic regulation of viral gene expression. The levels of Ast expression in single infected cells isolated from donors on suppressive antiretroviral therapy (ART) was investigated with a cell-associated RNA and DNA single-genome sequencing (CARD-SGS) assay adapted to detect Ast RNA. Additionally, a digital Polymerase Chain Reaction (PCR) approach was used to examine both sense and antisense HIV-1 gene expression. A median of ~5% of infected cells contained Ast RNA when measured by CARD-SGS. Ast expression levels ranged from 1-30 RNA copies/cell. Ast RNA measured using the digital reverse transcription PCR (RT-PCR) approach was detected in up to a median of 26% of infected peripheral blood mononuclear cells (PBMC). A similar fraction of infected cells contained sense env RNA as antisense Ast RNA in most donor samples. These findings confirm expression of HIV-1 Ast RNA in donors on ART. Detection of Ast RNA in unstimulated cells from donors on ART warrants the investigation of its bifunctional role as a coding and a regulatory RNA in vivo.	en_US
dc.format.extent	73 Pages	en_US
dc.identifier	doi:10.13016/m2kwtv-j7gv
dc.identifier.uri	http://hdl.handle.net/11603/24550
dc.language.iso	en_US	en_US
dc.relation.isAvailableAt	Hood College
dc.rights	CC0 1.0 Universal	en_US
dc.rights.uri	http://creativecommons.org/publicdomain/zero/1.0/	*
dc.title	INVESTIGATION OF HIV-1 ANTISENSE TRANSCRIPT EXPRESSION IN VIVO	en_US
dc.type	Text	en_US