A trial to convert the mouse susceptible to mouse hepahtis virus to resistant one by anti-sense RNA

反义RNA将小鼠肝炎病毒易感小鼠转为耐药小鼠的试验

基本信息

批准号：
62580037
负责人：
TAGUCHI Fumihiro
金额：
$ 1.41万
依托单位：
National Center of Neurology and Psychiatry
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62580037/
关键词：
anti-sense RNA mouse hepatitis virus RNAポリメレースマウス肝炎ウィルス anti-sense RNA

项目摘要

A big problem in the animal experiments using mice and rats is epidemic infections with highly disseminating and fatal viruses as mouse hepatitis virus (MHV) and sendai virus. Nowadays, most of aminal experiments are carried out by using specific-pathogen free (SPF)animals which are especially prone to be infected with such viruses. One of the approaches to overcome such problems is to establish the animals which are genetically resistant to virus infections. Thus, we have undertaken to convert susceptible mice to MHV to resistant ones by gene transfer. Anti-sense RNA is considered to spceifically inhibit the translation of mRNA (sense RNA) by hybridizing to sense RNA. In the virus replication as well, antisense RNA may inhibit virus replication by preventing the expression of a gene which is inevitable to virus replication. First of all, we tried to make susceptible cultured cells to MHV resistant by expressing antisense RNA in such cultured cells. We have prepared 54mer oligonucleotides identical to a part of MHV RNA polymerase gene and oligonucleotides complementary to them, anealed these oligouncleotides and make 20 to 36 tandem repeats of such sequence. These tandem repeats were inserted into experssion vectors with SV40 or metallothionein promoters to express anti-sense RNA and they were cotransfected on L cells with pSV2 neo. L cells grown in the presence of G418 were cloned and examined for the level of anti-sense RNA and resistance to MHV infection. As a whole, no correlation has bee observed between the anti-sense RNA level and resistance to MHV at present. We would like to further investigate on the relationship between anti-sense RNA and resistance to MHV infection to finally establish the system to make susceptible animals to resistant by anti-sense RNA.

在使用小鼠和大鼠的动物实验中，一个大问题是流行病感染，高度传播和致命病毒是小鼠肝炎病毒（MHV）和仙台病毒。如今，大多数氨基实验是通过使用特异性无病原（SPF）动物进行的，这些动物特别容易被这种病毒感染。克服此类问题的方法之一是建立对病毒感染具有遗传抗性的动物。因此，我们已承诺通过基因转移将易感小鼠转化为MHV。抗义RNA被认为可以通过杂交与感官RNA抑制mRNA（sense RNA）的翻译。在病毒复制中，反义RNA可以通过防止基因的表达来抑制病毒复制，而基因的表达是不可避免的病毒复制的。首先，我们试图通过在这种培养的细胞中表达反义RNA来使易感培养的细胞对MHV抗性。我们已经制备了54mer寡核苷酸与MHV RNA聚合酶基因的一部分相同，寡核苷酸与它们互补的寡核苷酸，并使这些寡蛋白酶溶解并进行了20至36个顺序重复。将这些串联重复插入与SV40或金属硫蛋白启动子以表达抗敏感RNA的载体中，并将其共转染在具有PSV2 NEO的L细胞上。克隆在G418存在下生长的L细胞，并检查抗敏感RNA的水平和对MHV感染的抗性。总体而言，目前抗敏感性RNA水平和对MHV的抗性之间没有观察到蜜蜂。我们想进一步研究抗敏感性RNA与对MHV感染的抗性之间的关系，以最终建立该系统，使易感动物具有抗Sensens RNA抗性。