title: Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells.
abstract: The envelope glycoprotein of human immunodeficiency virus (HIV) consists of an exterior glycoprotein (gp120) and a trans-membrane domain (gp41) and has an important role in viral entry into cells. HIV-1 entry has been validated as a clinically relevant anti-viral strategy for drug discovery. In the present work, several 2'-F substituted RNA aptamers that bind to the HIV-1(BaL) gp120 protein with nanomole affinity were isolated from a RNA library by the SELEX (Systematic Evolution of Ligands by EXponential enrichment) procedure. From two of these aptamers we created a series of new dual inhibitory function anti-gp120 aptamer-siRNA chimeras. The aptamers and aptamer-siRNA chimeras specifically bind to and are internalized into cells expressing HIV gp160. The Dicer-substrate siRNA delivered by the aptamers is functionally processed by Dicer, resulting in specific inhibition of HIV-1 replication and infectivity in cultured CEM T-cells and primary blood mononuclear cells (PBMCs). Moreover, we have introduced a 'sticky' sequence onto a chemically synthesized aptamer which facilitates attachment of the Dicer substrate siRNAs for potential multiplexing. Our results provide a set of novel inhibitory agents for blocking HIV replication and further validate the use of aptamers for delivery of Dicer substrate siRNAs.
authors.affiliation: Division of Molecular Biology, Beckman Research Institute of City of Hope, City of Hope, Duarte, CA 91010, USA.
authors.forename: Jiehua
authors.initials: J
authors.lastname: Zhou
authors.forename: Piotr
authors.initials: P
authors.lastname: Swiderski
authors.forename: Haitang
authors.initials: H
authors.lastname: Li
authors.forename: Jane
authors.initials: J
authors.lastname: Zhang
authors.forename: C Preston
authors.initials: CP
authors.lastname: Neff
authors.forename: Ramesh
authors.initials: R
authors.lastname: Akkina
authors.forename: John J
authors.initials: JJ
authors.lastname: Rossi
mesh_terms: D000818:Animals; D019380:Anti-HIV Agents / Q000737:chemistry* / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism; D001483:Base Sequence; D002478:Cells, Cultured; D015699:HIV Envelope Protein gp120 / Q000037:antagonists & inhibitors* / Q000235:genetics; D019302:HIV Envelope Protein gp160 / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000378:metabolism; D043244:Ribonuclease III / Q000378:metabolism; D052156:SELEX Aptamer Technique; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D019380:Anti-HIV Agents; D052157:Aptamers, Nucleotide; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D007370:Interferon Type I; D034741:RNA, Small Interfering; C514793:gp120 protein, Human immunodeficiency virus 1; C514797:gp160 protein, Human immunodeficiency virus 1; D043244:Ribonuclease III