权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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，一个大问题是高度传播和致命病毒的流行性感染，如小鼠肝炎病毒（MHV）和仙台病毒。目前，大多数动物实验都是使用SPF级动物进行的，而SPF级动物是最容易感染此类病毒的动物。克服这些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，我们已经着手通过基因转移将MHV易感小鼠转化为耐药小鼠。反义RNA被认为是通过与有义RNA杂交来特异性抑制mRNA（有义RNA）的翻译。同样在病毒复制中，反义RNA可以通过阻止病毒复制所不可避免的基因的表达来抑制病毒复制。首先，我们试图通过在这样的培养细胞中表达反义RNA来使对MHV敏感的培养细胞具有抗性。我们制备了与MHV RNA聚合酶基因的一部分相同的54聚体寡核苷酸和与之互补的寡核苷酸，并将这些寡核苷酸退火，使该序列形成20 ~ 36个串联重复。将这些串联重复序列分别插入到含有SV 40或金属硫蛋白启动子的表达载体中，表达反义RNA，并与pSV 2 neo共转染L细胞。将在G418存在下生长的L细胞克隆并检查反义RNA的水平和对MHV感染的抗性。总体而言，目前尚未观察到反义RNA水平与MHV抗性之间的相关性。我们希望进一步研究反义RNA与抗MHV感染之间的关系，最终建立通过反义RNA使易感动物产生抗MHV感染的系统。