Development of the gene expression inhibition system using ribozyme

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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.