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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Effective inhibition in animals of viral pathogenesis by a ribozyme derived from RNase P catalytic RNA
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Effective inhibition in animals of viral pathogenesis by a ribozyme derived from RNase P catalytic RNA

机译：源自RNase P催化RNA的核酶有效抑制动物的病毒发病机制。

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摘要

A functional RNase P ribozyme (M1GS RNA) was constructed to target the overlapping mRNA region of two murine cytomegalovirus (MCMV) capsid proteins essential for viral replication: the assembly protein (mAP) and M80. The customized ribozyme efficiently cleaved the target mRNA sequence in vitro. Moreover, 80% reduction in the expression of mAP and M80 and a 2,000-fold reduction in viral growth were observed in cells expressing the ribozyme. In contrast, there was no significant reduction in viral gene expression and growth in cells that either did not express the ribozyme or produced a "disabled" ribozyme carrying mutations that abolished its catalytic activity. When the ribozyme-expressing constructs were delivered into MCMV-infected SCID mice via a modified "hydrodynamic transfec-tion" procedure, expression of ribozymes was observed in the livers and spleens. Compared with the control animals that did not receive any M1GS constructs or received the disabled ribozyme construct, animals receiving the functional ribozyme construct exhibited a significant reduction of viral gene expression and infection. Viral titers in the spleens, livers, lungs, and salivary glands of the functional ribozyme-treated SCID mice at 21 days after infection were 200- to 2,000-fold lower than those in the control animals. Moreover, survival of the infected animals significantly improved upon receiving the functional ribozyme construct. Our study examines the use of M1GS ribozymes for inhibition of gene expression in animals and demonstrates the utility of RNase P ribozymes for gene targeting applications in vivo.
机译：构建功能性RNase P核酶（M1GS RNA），以靶向两种对病毒复制必不可少的鼠巨细胞病毒（MCMV）衣壳蛋白的重叠mRNA区域：装配蛋白（mAP）和M80。定制的核酶在体外有效切割靶标mRNA序列。此外，在表达核酶的细胞中，mAP和M80的表达降低了80％，病毒生长降低了2000倍。相反，在不表达核酶或产生携带突变而废除了其催化活性的“残疾”核酶的细胞中，病毒基因的表达和生长没有显着减少。当通过改进的“流体动力转化”方法将表达核酶的构建体递送至感染了MCMV的SCID小鼠中时，在肝脏和脾脏中观察到了核酶的表达。与不接受任何M1GS构建体或接受禁用的核酶构建体的对照动物相比，接受功能性核酶构建体的动物表现出病毒基因表达和感染的显着降低。感染后第21天，功能核酶处理的SCID小鼠的脾脏，肝脏，肺脏和唾液腺中的病毒滴度比对照动物低200-2,000倍。此外，在接受功能性核酶构建体后，感染动物的存活率显着提高。我们的研究检查了M1GS核酶在动物中抑制基因表达的用途，并证明了RNase P核酶在体内基因靶向应用中的实用性。

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