TY - JOUR
T1 - Delivery of double-stranded DNA thioaptamers into HIV-1 infected cells for antiviral activity
AU - Ferguson, Monique R.
AU - Rojo, Daniel R.
AU - Somasunderam, Anoma
AU - Thiviyanathan, Varatharasa
AU - Ridley, Bettye D.
AU - Yang, Xianbin
AU - Gorenstein, David G.
N1 - Funding Information:
We thank the NIH AIDS Research and Reference Reagent Program for providing the HIV-1 SF162-R5 and U373-MAGI-CCR5 cells. This work was supported by NIH Grants R21 AI058194-01A2 (M.R.F.), U01-A1054827, ES06676 and A127744 (D.G.G.) and by the Welch Foundation (H-1296, to D.G.G.).
PY - 2006/6/9
Y1 - 2006/6/9
N2 - 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.
AB - 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.
KW - HIV
KW - Liposomes
KW - Oligonucleotide
KW - RNase H
KW - RT inhibitors
KW - Thioaptamer
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U2 - 10.1016/j.bbrc.2006.03.201
DO - 10.1016/j.bbrc.2006.03.201
M3 - Article
C2 - 16631118
AN - SCOPUS:33646125429
SN - 0006-291X
VL - 344
SP - 792
EP - 797
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -