Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells.
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.
Zhou
Jiehua
J
Division of Molecular Biology, Beckman Research Institute of City of Hope, City of Hope, Duarte, CA 91010, USA.
Swiderski
Piotr
P
Li
Haitang
H
Zhang
Jane
J
Neff
C Preston
CP
Akkina
Ramesh
R
Rossi
John J
JJ
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
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