Potent and selective gene inhibition using antisense oligodeoxynucleotides.
The development of antisense technology as a generally useful tool relies on the use of potent agents and the utilization of many controls in experiments. Here we describe our experience using oligodeoxynucleotides (ODNs) containing C-5 propynyl pyrimidine and phosphorothioate modifications as broadly applicable gene inhibition agents in cell culture. Methods include selection of antisense sequences, synthesis and purification of ODNs, choice of controls, delivery methods (microinjection, cationic lipid transfection, and electroporation), and analysis of gene inhibition.
Flanagan
W M
WM
Gilead Sciences, Foster City, CA 94404, USA.
Wagner
R W
RW
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000235:genetics; D002412:Cations; D002460:Cell Line; D002522:Chlorocebus aethiops; D018274:Electroporation; D005786:Gene Expression Regulation / Q000187:drug effects*; D050356:Lipid Metabolism; D008845:Microinjections; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D012313:RNA / Q000032:analysis; D012260:Ribonucleases / Q000378:metabolism; D013873:Thionucleotides
D002412:Cations; D016376:Oligonucleotides, Antisense; D013873:Thionucleotides; D012313:RNA; D016203:CDC2 Protein Kinase; D012260:Ribonucleases