GENERATION OF MOUSE MODELS FOR HUMAN GENETIC DISEASESTHROUGH GENE TARGETING

通过基因靶向生成人类遗传疾病小鼠模型

基本信息

批准号：
07457040
负责人：
SHIMADA Kazunori
金额：
$ 4.86万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

1) A replacement vector convenient for introducing subtle mutations into various mouse genes has been developed using as a model system, the mouse transthyretin-encoding gene (ttr) and mouse embryonal carcinoma F9 cells. The vector consists of part of ttr carrying a subtle mutation in its second exon, and a cassette of the neomycin-resistance (neo) - and herpes simplex virus thymidine kinase (HSV-tk) -encoding genes flanked with a 3-kb duplication of mostly the second intron of ttr.2) To study gene expression in undifferentiated mouse embryonal carcinoma F9 cell, we prepraed 2,132 expressed sequence tags (ESTs) and found that 1,416 match known gene and/or protein sequences. We tried to develop a system for characterizing ESTs matching no known genes. We also isolated 17 cDNA clones corresponding to mRNAs induced rapidly during retinoic acid-mediated F9 cell differentiation and characterized two of them, named rael and rae28, in this study.3) To study the role of the rae28gene in mouse development, we generated rae28-deficient mice by gene targeting in embryonic stem cells. To our surprise, the homozygous rae28-knock out mice carried all the phenotypes noted in the human congenital disorder CATCH-22 syndrome. We found that the anterior boundaries of Hoxa-3, a-4, a-5, b-3, b-4 and d-4 expression are shifted in the rostral direction in the paraxial mesoderms of the rae28-/- homozygous embryos, and those of Hoxb-3 and b-4 expression are also similarly altered in the rhombomeres and/or pharyngeal arches. These altered Hox codes were presumed to be correlated with the posterior skeletal transformations and neural crest defects observed in the rae28-/- homozygous mice. These results indicate that the rae28 gene is involved in the regulation of Hox gene expression and segment specification during paraxial mesoderm and neural crest development.4) We continued studies on mouse models for familial amyloidotic polyneuropathy.

1）替换矢量方便用于将细微突变引入各种小鼠基因的方便，已作为模型系统，小鼠经甲状腺素蛋白编码基因（TTR）和小鼠胚胎癌F9细胞开发。载体由TTR组成的一部分在其第二个外显子中携带微妙的突变，以及新霉素抗性（NEO）的盒式和单纯疱疹病毒胸苷激酶（HSV-TK）的疱疹，将基因赋予基因，并以3-kb的范围射击了TTR的第二个crone forne fene ferc and gene in n embry fene in gene in n embry formand.2细胞，我们准备了2,132个表达的序列标签（EST），发现1,416个匹配已知基因和/或蛋白质序列。我们试图开发一个用于表征没有已知基因的ESTS的系统。我们还分离了在视黄酸介导的F9细胞分化过程中迅速诱导的MRNA诱导的17个cDNA克隆，并在这项研究中表征了其中两个，称为Rael和Rae28。令我们惊讶的是，纯合子RAE28敲除小鼠携带了人类先天性疾病捕获22综合征中注意到的所有表型。 We found that the anterior boundaries of Hoxa-3, a-4, a-5, b-3, b-4 and d-4 expression are shifted in the rostral direction in the paraxial mesoderms of the rae28-/- homozygous embryos, and those of Hoxb-3 and b-4 expression are also similarly altered in the rhombomeres and/or pharyngeal arches.假定这些改变的HOX代码与RAE28 - / - 纯合小鼠中观察到的后骨骼转化和神经rest缺陷相关。这些结果表明，RAE28基因参与了近期中胚层和神经rest发育过程中HOX基因表达和片段规范的调节。4）我们继续研究小鼠模型的家族性淀粉样蛋白多神经病。