title: Characterization of an HIV-targeted transcriptional gene-silencing RNA in primary cells.
abstract: Small antisense RNAs targeted to the HIV-1 promoter have been shown to remodel the surrounding chromatin to a state unfavorable for transcriptional activation, yet transcriptional gene silencing (TGS) of HIV-1 has, to date, not been shown in primary human cells. We demonstrate here that TGS can reduce viral transcription in primary human CD4(+) T cells; however, increasing viral burden results in the loss of this antiviral effect. This observation suggests a critical level at which viral RNA can dilute out effective targeting by TGS-based RNAs. Furthermore, studies into off-target effects have identified a potential interaction between the small nucleolar RNA pathway and the TGS-based antisense RNA, resulting in activation of p53. Although not overtly toxic to primary cells, this represents a novel interaction between antisense RNAs and a cellular pathway that should be considered when pursuing small antisense RNA-based therapeutics.
authors: Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors: Anne-Marie W
authors: AM
authors: Turner
authors: Amanda M
authors: AM
authors: Ackley
authors: Michael A
authors: MA
authors: Matrone
authors: Kevin V
authors: KV
authors: Morris
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism / Q000821:virology*; D002843:Chromatin / Q000378:metabolism; D020868:Gene Silencing; D005822:Genetic Vectors / Q000378:metabolism; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000235:genetics; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D002843:Chromatin; D016372:RNA, Antisense; D016159:Tumor Suppressor Protein p53