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外部引导序列(EGSs)是一类与mRNA靶序列互补并能引导核酶P切割靶mRNA的小分子RNA。本实验构建稳定表达UL49基因的HeLa细胞系,设计合成了针对于人巨细胞病毒(HCMV)UL49基因的12ntDNA性质的EGS1386,通过转染稳定表达UL49基因的细胞系,荧光定量PCR和Western blotting检测细胞内目的基因UL49的表达情况。结果显示在DNA-EGS1386作用下UL49基因的表达量降低了50%,表明DNA-EGS1386可以有效引导人的核酶P切割目标mRNA。因此,DNA-EGS可以发展成为一种新的基因沉默技术和潜在的抗病毒试剂。
External leader sequences (EGSs) are a class of small RNAs that are complementary to mRNA target sequences and can direct ribozyme P to cleave target mRNAs. In this study, a HeLa cell line stably expressing UL49 gene was constructed and designed to synthesize a 12 ntDNA-specific EGS1386 directed against UL49 gene of human cytomegalovirus (HCMV). The transfected cell line stably expressing UL49 gene was detected by quantitative PCR and Western blotting UL49 target gene expression in the cell. The results showed that under the action of DNA-EGS1386, the expression of UL49 gene was reduced by 50%, indicating that DNA-EGS1386 can effectively guide human ribozyme P to cleave the target mRNA. Therefore, DNA-EGS can develop into a new gene silencing technology and a potential anti-viral agent.