DNA-EGS1386胞内诱导核酶P抑制人巨细胞病毒UL49基因的表达

DNA-EGS1386 in cells induced RNase P inhibits the expression of human cytomegalovirus UL49 gene

摘要:

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12 nt DNA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。

Abstract:

External Guide Sequences (EGSs) represents a novel nucleic acid based gene interference approach to modulate gene expression. They are oligonucleotides that consist of a sequence complementary to a target mRNA and recruit intracellular RNase P for specific degradation of the target RNA. DNA-based EGS1386 with a size of 12 nt was chemically synthesized to target the mRNA coding for the UL49 gene of human cytomegalovirus (HCMV). The DNA-based EGS1386 molecule efficiently directed human RNase P to cleave the target mRNA sequence in vitro. A reduction of more than 50% in the levels of UL49 expression was observed in human cells treated with the DNA-based EGS1386 targeted UL49 assayed by fluorescent quantization PCR and Western blotting. This results showed that the DNA-EGS1386 can effectively guide the RNase P cut the target mRNA. Therefore, DNA-EGS can develop into a new gene silencing technology and potential of the anti-viral reagents.