RNase P体外靶定切割人巨细胞病毒UL54 mRNA片段研究

In vitro targeted cleavage of human cytomegalovirus UL54 mRNA segments by RNase P

摘要: 目的:观察大肠埃希菌RNase P催化亚基M1 RNA在细胞外水平上对人巨细胞病毒(human cytomegalovirus,HCMV)DNA聚合酶UL54 mRNA的切割效果,探讨核酶作为新型抗病毒制剂的应用前景。方法:针对HCMVUL54 mRNA设计特异性的外部引导序列(external guide sequences,EGSs)EGS-T6,观察其引导M1 RNA特异的细胞外切割活性。结果:在EGS-T6引导下,大肠埃希菌RNase P催化亚基M1 RNA在细胞外可特异地对靶mRNA进行有效切割。结论:EGS-T6具备引导M1 RNA对UL54 mRNA特异切割的能力,可发展成一种新型抗病毒制剂。

Abstract: Objective:To study the cleavage effect of M1 RNA,catalytic subunit of RNase P from Escherichia colion on UL54 mRNA encoding major DNA polymerase of human cytomegalovirus in vitro,and to explore the prospects of ribozyme as a novel antiviral agent.Methods:An external guiding sequence T6 specific to HCMV UL54 mRNA was designed and synthesized;specific cleavage activity of M1RNA guided by T6 was observed in vitro.Results:With the guidance of EGS-T6,M1 RNA from Escherichia coli can block the expression of HCMV UL54 mRNA in vitro.Conclusions:EGS-T6 may fulfill the specific cleavage and be developed as a novel antiviral agent.