Delivery of double-stranded DNA thioaptamers into HIV-1 infected cells for antiviral activity.
Oligonucleotide agents (ODN) are emerging as attractive alternatives to chemical drugs. However, the clinical use of ODNs as therapeutics has been hindered by their susceptibility to degradation by cellular enzymes and their limited ability to penetrate intact cells. We have used various liposome-mediated transfection agents, for the in vitro delivery of DNA thioaptamers into U373-MAGI-CCR5 cells. Our lead thioaptamer, R12-2, targets the RNase H domain of the HIV-1 reverse transcriptase (RT) and inhibits viral infection in U373-MAGI-CCR5 cells. R12-2, a 62-base-pair, double-stranded DNA molecule with a monothio-phosphate modified backbone, was selected through a novel combinatorial selection method. We studied the use of oligofectamine (OF), TFX-20, Transmessenger (TM), and Gene Jammer (GJ) for transfection of the thio-modified DNA aptamers. OF-transfected U373-MAGI-CCR5 cells resulted in 68% inhibition of HIV infection in the treated cells compared to the untreated control. Inhibition was observed in a dose-dependent manner with maximal inhibition of 83%. In this report, we demonstrate that monothioate-modified DNA duplex oligonucleotides can be efficiently delivered into cells by liposome-based transfection agents to inhibit HIV replication.
Ferguson
Monique R
MR
Division of Infectious Diseases, Department of Internal Medicine, The University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA. mrfergus@utmb.edu
Rojo
Daniel R
DR
Somasunderam
Anoma
A
Thiviyanathan
Varatharasa
V
Ridley
Bettye D
BD
Yang
Xianbin
X
Gorenstein
David G
DG
D000998:Antiviral Agents / Q000008:administration & dosage; D052157:Aptamers, Nucleotide / Q000008:administration & dosage* / Q000235:genetics*; D001254:Astrocytoma / Q000201:enzymology / Q000235:genetics* / Q000821:virology*; D045744:Cell Line, Tumor; D018390:Gene Targeting / Q000379:methods; D015316:Genetic Therapy / Q000379:methods; D015658:HIV Infections / Q000235:genetics / Q000628:therapy; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D016914:Ribonuclease H / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods
D000998:Antiviral Agents; D052157:Aptamers, Nucleotide; D016914:Ribonuclease H