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Development of the gene expression inhibition system using ribozyme

使用核酶开发基因表达抑制系统

基本信息

批准号：
14560240
负责人：
HISAMATSU Shin
金额：
$ 1.79万
依托单位：
Azabu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560240/
关键词：
ribozyme gene expression inhibition pseudorabies virus ブタウイルス

项目摘要

Ribozymes are capable of catalyzing sequence specific cleavate of RNA. Therefore, if the function of this ribozyme is used, gene expression may be controllable by the mRNA level. In this study, ribozymes were designed to RNAs of Pseudorabies virus which is the cause virus of the Aujeszky's disease of a pig. And, because the pig of the virus resistance using ribozyme might be producted, it planned introducing genes of these ribozymes for RNA of IE180 and EPO gene into a pig. Consequently, the designed ribozyme was able to cleave the full-length RNA of IE180 and EPO gene. Moreover, when this ribozyme gene was introduced into the pig cultivation cell and Pseudorabies virus resistance was investigated, a certain level of resistance was shown. However, it thought that virus resistance with stronger using RNAi whose gene expression control is enabled effectively was expectable rather than the ribozyme, and planned experimenting in the virus resistance using RNAi. By the way, GC content of IE180 gene as target is very high, about 80%, and its GC content of an EPO gene is also high. Since it is thought that transcripts of the high GC content genes have taken high order structure and the siRNA itself to introduce may take high order structure too, it is expected that it is difficult to be these RNA to target. Therefore, it is thought that the important knowledge about production of virus tolerance animal is acquired by developing energetic research to this target.

核酶能够催化RNA的序列特异性裂解。因此，如果利用这种核酶的功能，基因的表达可能受到mRNA水平的控制。在这项研究中，核糖酶被设计为猪瘟的致病病毒伪狂犬病毒的rna。并且，由于利用核酶对病毒产生抗性的猪可能被生产出来，因此计划将这些核酶的基因用于IE180和EPO基因的RNA导入猪体内。因此，所设计的核酶能够切割IE180和EPO基因的全长RNA。此外，将该核酶基因引入猪培养细胞，并对假狂犬病毒进行抗性研究，显示出一定程度的抗性。然而，他们认为使用RNAi而不是核酶更能有效地控制基因表达，从而产生更强的病毒抗性是可以预期的，并计划使用RNAi进行病毒抗性实验。顺便说一下，IE180基因作为靶基因的GC含量非常高，约为80%，其一个EPO基因的GC含量也很高。由于认为高GC含量基因的转录本具有高阶结构，并且引入的siRNA本身也可能具有高阶结构，因此预计这些RNA很难被靶向。因此，通过对该靶点的深入研究，可以获得生产耐病毒动物的重要知识。