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

使用核酶开发猪病毒控制系统

• 批准号: 11660329
• 负责人: KASHIWAZAKI Naomi
• 金额: $ 1.86万
• 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: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11660329/
• 关键词: ribozyme; virus; pig; pseudorabies virus; ウイルス防除; PRV

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.

利用核酶建立猪病毒控制系统核酶能够催化RNA的序列特异性切割，因此，它们已成为通过切割RNA来人工控制基因表达的潜在工具。为培育抗伪狂犬病猪，设计了针对伪狂犬病病毒（PRV）即早180基因mRNA的发夹状核酶，并利用猪培养细胞检测了核酶的活性。将转录RNA聚合酶II（CMV增强子和启动子）或III（tRNA）的启动子的riobzyme基因导入猪培养细胞。用RNA聚合酶Ⅱ转录核酶基因的细胞感染PRV，感染后第1天细胞出现CPE。然后CPE在三天内遍布整个培养物。这一结果与未导入核酶基因的猪培养细胞中的结果基本一致。然而，用RNA聚合酶III转录核酶基因的培养细胞显示出对PRV感染的抗性，表明CPE在感染后6天内没有在培养物中扩散。

项目成果

Nishino,Y. et al: "Borna disease virus infection in domestic cats:Evaluation by RNA and antibody detection."J.Vet.Med.Sci.. 61・10. 1167-1170 (1999)

Nishino, Y.等人：“家猫中的博尔纳病病毒感染：通过RNA和抗体检测进行评估。”J.Vet.Med.Sci. 61・10（1999）。