Enhanced gene silencing by the application of multiple specific small interfering RNAs.
Small interfering RNA duplexes (siRNA) induce gene silencing in various eukaryotic cells, although usually in an incomplete manner. Using chemically synthesized siRNAs targeting the HIV-1 co-receptor CXCR4 or the apoptosis-inducing Fas-ligand (FasL), co-transfection of cells with two or more siRNA duplexes targeting different sites on the same mRNA resulted in an enhanced gene silencing compared with each single siRNA. This was shown in the down-regulation of protein and mRNA expression, and functionally in the inhibition of CXCR4-mediated HIV infection and of FasL-mediated cell apoptosis. Transfection efficiency determined for the FasL-specific siRNAs was dose-dependent and varied among the siRNAs tested, but was not the main reason for the enhanced gene silencing.
Ji
Jingmin
J
Institute for Medical Microbiology, University of Basel, Petersplatz 10, 4003 Basel, Switzerland.
Wernli
Marion
M
Klimkait
Thomas
T
Erb
Peter
P
D017209:Apoptosis; D001483:Base Sequence; D002460:Cell Line; D015536:Down-Regulation; D053222:Fas Ligand Protein; D020868:Gene Silencing; D015497:HIV-1 / Q000472:pathogenicity; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000138:chemical synthesis / Q000235:genetics*; D019718:Receptors, CXCR4 / Q000235:genetics; D014162:Transfection
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D034741:RNA, Small Interfering; D019718:Receptors, CXCR4