Generation of hypomorphic model mice utilizing an mRNA instability element

利用 mRNA 不稳定元件生成亚效型模型小鼠

基本信息

批准号：
17500286
负责人：
MORI Masayuki
金额：
$ 2.24万
依托单位：
SHINSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17500286/
关键词：
hypomorph model mouse mRNA instability element 遺伝子ゲノム動物

项目摘要

We identified the genes associated with cataract formation in Shumiya cataract rat (SCR) by positional cloning. Mutations of the gene for lanosterol synthase (Lss) were identified. Cataract onset in SCR was uniquely regulated by a specific combination of different mutant (Lss^l) and polymorphic alleles (Lss^s) on the Lss locus. Lss^s was a hypomorphic allele with a G to A nucleotide substitution in exon 4. The G to A nucleotide substitution also caused instability of the Lss mRNA transcripts.When a luciferase reporter gene was linked to the exon 4 sequence of Lss^s, which contain the mRNA instability element and expressed in the COS cells, luciferase activity was significantly reduced. This result indicated that the mRNA instability element functions regardless of the gene, in which it is located. We then examined the possibility that the change of an exonic splicing enhancer (ESE) element by the G to A substitution is the cause of instability of the Lss^s mRNA precursor. Analysis by the computer program ESEfinder identified a novel SRp40 binding motif, with a score of 4.15, in the Lss^s allele. SRp40 has previously been studied for its role in alternative splicing and has been shown to select both proximal and distal 5' splice sites in a substrate-specific manner. It was suggested that the Lss^s mRNA precursor becomes unstable if SRp40 splicing factor is bound to irrelevant sites in the precursor.

我们通过位置克隆确定了与白内障大鼠（SCR）中白内障形成相关的基因。鉴定了羊毛醇合酶基因的突变（LSS）。 SCR中的白内障发作受到LSS基因座上不同突变体（LSS^L）和多态性等位基因（LSS^s）的特定组合的独特调节。 Lss^s was a hypomorphic allele with a G to A nucleotide substitution in exon 4. The G to A nucleotide substitution also caused instability of the Lss mRNA transcripts.When a luciferase reporter gene was linked to the exon 4 sequence of Lss^s, which contain the mRNA instability element and expressed in the COS cells, luciferase activity was significantly reduced.该结果表明mRNA不稳定性元件的功能不管其所在的基因如何。然后，我们研究了G替换的外显子剪接增强子（ESE）元素的变化是LSS s mRNA前体不稳定性的原因。计算机程序ESEFINDER的分析确定了一个新型的SRP40结合基序，分数为4.15，在LSS等位基因中。先前已经研究了SRP40在替代剪接中的作用，并已证明可以以底物特异性方式选择近端和远端5'剪接位点。有人提出，如果SRP40剪接因子与前体中的位点无关，则LSS的mRNA前体变得不稳定。

项目成果

期刊论文数量（28）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A locus for eosinophilia in the MES rat is on Chromosome 19

DOI：
10.1007/s00335-004-2454-5
发表时间：
2005-07-01
期刊：
MAMMALIAN GENOME
影响因子：
2.5
作者：
Li, GX;Guo, ZJ;Mori, M
通讯作者：
Mori, M

Secreted Frizzled-Related Protein 4 as a negative regulator of bone formation and a candidate gene affecting peak bone mineral density in mice

分泌性卷曲相关蛋白 4 作为骨形成的负调节因子和影响小鼠峰值骨矿物质密度的候选基因

DOI：
发表时间：
2006
期刊：
Journal of Bone and Mineral Research 21・11
影响因子：
0
作者：
Nakanishi R.;Shimizu M.;Mori M.;Akiyama H.;Otsuki B.;Okudaira S.;Hashimoto M.;Higuchi K.;Tsuboyama T.;Nakamura T.
通讯作者：
Nakamura T.

Reduced coenzyme Q10 supplementation decelerates senescence in SAMP1 mice