Zhuang Fengfeng, Liu Yi-Hsin
Center for Craniofacial Molecular Biology and Graduate Program in Cranifacial Biology, School of Dentistry, University of Southern California, Los Angeles, CA, USA.
Methods Mol Biol. 2006;342:181-7. doi: 10.1385/1-59745-123-1:181.
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.
4年前Tushl等人首次证明小干扰RNA（siRNA）后，它革新了培养的哺乳动物细胞中的基因沉默技术。为了规避在鉴定能够抑制靶基因表达的独特siRNA双链体时的成本和不便，我们设计了一种基于报告基因的系统来测试所选siRNA的敲低效率。我们证明，这种基于荧光素酶的siRNA测试系统可用于评估直接转染的siRNA双链体或从慢病毒载体表达的siRNA的敲低效率。
