Generation of knockout mice for Nijmegen breakage syndrome gene, NBS1.

奈梅亨断裂综合征基因 NBS1 基因敲除小鼠的产生。

• 批准号: 11138237
• 负责人: MATSUURA Shinya
• 金额: $ 2.69万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas (A)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11138237/
• 关键词: Nijmegen syndrome; Cancer prone disease; Ataxia-telangiectasia; Rodiation sensitivity; Gene targeting; Model mouse; Embryonic lethal; Nbs1; Nbs1

Nijmegen breakage syndrome is an autosomal recessive disorder characterized by combined immunodeficiency, predisposition to cancer, microcephaly, and growth retardation. Cells from Nijmegen breakage syndrome display chromosomal instability, radiation sensitivity, and radioresistant DNA synthesis. An Nbs1 targeting vector was constructed and was introduced into mouse ES cells by electroporation. G418 resistant ES cell clones were selected and were screened for homologous recombination by Southern blot analysis. Out of 234 clones, 3 clones showed correct recombination and were used for blastocyst injection. Chimeric mice were back crossed with C57BL/6, and F1 mice were generated. PCR analysis demonstrated germ line transmission of the Nbs 1 mutant allele. Although the Nbs1 heterozygous mice were normal and fertile, Nbs1 null mice could not be obtained so far. The result suggested that the disruption of Nbs1 could cause embryonic lethality, indicating that Nbs1 protein complex is essential for viability. We are setting up timed matings to investigate the cause of embryonic lethality, and also trying to establish Nbs1 nullizygous fibroblasts to define further function of the Nbs1 gene.

Nijmegen Breakage综合征是一种常染色体隐性障碍，其特征是免疫缺陷，癌症，小头畸形和生长迟缓的易感性。 Nijmegen破裂综合征的细胞表现出染色体不稳定性，辐射敏感性和放射性DNA合成。构建了NBS1靶向载体，并通过电穿孔引入小鼠ES细胞中。选择G418抗性ES细胞克隆，并通过Southern印迹分析筛选出同源重组。在234个克隆中，有3个克隆表现出正确的重组，并用于胚泡注射。将嵌合小鼠与C57BL/6交叉，并产生F1小鼠。 PCR分析证明了NBS 1突变等位基因的生殖线传递。尽管NBS1杂合小鼠正常且肥沃，但到目前为止无法获得NBS1无效小鼠。结果表明，NBS1的破坏可能引起胚胎致死性，表明NBS1蛋白复合物对于生存力至关重要。我们正在设置定时整体以研究胚胎致死性的原因，并试图建立NBS1无曲纤维成纤维细胞来定义NBS1基因的进一步功能。

项目成果

Morishima,K., et al.: "A polymorphic CA repeat marker at the human 27-KD calbindin(CALB1) locus"J.Hum.Genet.. 44. 414-415 (1999)

Morishima,K., et al.：“人 27-KD 钙结合蛋白 (CALB1) 基因座的多态性 CA 重复标记”J.Hum.Genet.. 44. 414-415 (1999)

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Nakamura, A.、Matsuura, S.、Tauchi, H.、Hanada, R.、Ohashi, H.、Hasekawa, T.、Honda, K.、Masuno, M.、Imaizumi, K.、Sugita, K.、