Usefulness of the luciferase reporter system to test the efficacy of siRNA.
Small interefence RNA (siRNA) has revamped the technology of gene silencing in cultured mammalian cells after its first demonstration by Tushl et al. 4 yr ago. To circumvent the cost and the inconvenience in identifying a unique siRNA duplex that can quench target gene expression, we devised a reporter-based system to test knockdown efficiency of selected siRNAs. We demonstrated that this luciferase-based siRNA testing system can be used to evaluate the knockdown efficiency of a directly transfected siRNA duplex or an siRNA expressed from a lentiviral vector.
Zhuang
Fengfeng
F
Center for Craniofacial Molecular Biology and Graduate Program in Cranifacial Biology, School of Dentistry, University of Southern California, Los Angeles, CA, USA.
Liu
Yi-Hsin
YH
D000818:Animals; D002460:Cell Line; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D016086:Lentivirus; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D032446:Myoblasts / Q000378:metabolism; D010957:Plasmids; D034741:RNA, Small Interfering / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D014162:Transfection
D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D008156:Luciferases