Development of the virus-control system in pigs using the ribozyme

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Development of the virus-control system in pigs using the ribozymeRibozymes are capable of catalyzing sequence specific cleavage of RNA.Therefore they have been emerging as potential tools to artificially control gene expression through cleavage of RNAs. For breeding of the pig which has resistance to Aujeszkey's disease caused by the infection of pseudorabies virus (PRV), we designed hairpin ribozyme for mRNA of immediate early 180 gene of the virus and examined the activity of ribozyme using pig cultured cell. The riobzyme gene transcribed the promoter of RNA polymerase II (CMV enhancer and promoter) or III (tRNA) was introduced to the pig cultured cells. The cultured cell, in which the ribozyme gene was transcribed by RNA polymerase II was infected with PRV, CPE was observed in cultured cell on the first day after infection. And then CPE spread over the whole culture within three days. This result was almost the same as that in the pig cultured cell which ribozyme gene was not introduced. However, the cultured cell, in which the ribozyme gene was transcribed by RNA polymerase III, revealed the resistance to PRV infection showing that CPE didn't spread over the whloe culture by six days after infection.