针对HCV基因组中较为保守的区域-5'UTR,设计一段GS引导序列,并与大肠杆菌RNase P的催化亚基-M1RNA的3'末端共价结合,构建序列特异性M1GS核酶-M1GS-HCV/C20。体外实验证实,所构建的人工核酶对HCV 5'UTR具有明显的靶向切割活性,且这种切割发生于靶序列的特定位点。本研究将为进一步阐明该核酶在胞内的活性、乃至动物模型内评价其抗病毒效果提供实验材料,从而为新型抗HCV药物及反义基因治疗的研究奠定基础。
In this study, a sequence-specific M1GS ribozyme (M1GS-HCV/C20) has been successfully constructed by covalently linking an oligonucleotide (guide sequence, GS) to the 3' terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The engineered ribozyme is targeted to the most conservative sequence (5'UTR) of HCV genome, and can effectively cleave the substrate RNA segment in vitro. Undoubtly, the M1GS-HCV/C20 we got would be a useful experimental material to futher study its cleavage activity in vivo, and can be even used for evaluating its anti-viral effect in the animal model. It was believed that this study would markedly facilitate the research of a general gene targeting agent for anti-HCV applications, and layed the foundation for developing a new nucleic acid drug and a novel strategy of anti-HCV therapy.
张文军,宁容,张欣,李红枝. HCV特异性M1GS核酶的构建及体外切割活性研究[J]. 中国生物工程杂志, 2008, 28(9): 99-103.
. Study on the in vitro cleavage activity of an artificial HCV-specific M1GS ibozyme. China Biotechnology, 2008, 28(9): 99-103.
https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2008/V28/I9/99
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