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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Spontaneous Irs1 passenger mutation linked to a gene-targeted SerpinB2 allele
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Spontaneous Irs1 passenger mutation linked to a gene-targeted SerpinB2 allele

机译:自发性Irs1乘客突变与靶向基因的SerpinB2等位基因相关

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摘要

In characterizing mice with targeted disruption of the SerpinB2 gene, we observed animals that were small at birth with delayed growth and decreased life expectancy. Although this phenotype cosegregated with homozygosity for the inactive SerpinB2 allele, analysis of homozygous SerpinB2-deficient mice derived from two additional independent embryonic stem (ES) cell clones exhibited no growth abnormalities. Examination of additional progeny from the original 5erpinB2-deficient line revealed recombination between the small phenotype (smla) and the SerpinB2 locus. The locus responsible for smla was mapped to a 2.78-Mb interval approximately 30 Mb proximal to SerpinB2, bounded by markers D1Mit382 and D1Mit216. Sequencing of Irs1 identified a nonsense mutation at serine 57 (SS7X), resulting in complete loss of IRS1 protein expression. Analysis of ES cell DNA suggests that the S57X Irs1 mutation arose spontaneously in an ES cell subclone during cell culture. Although the smla phenotype is similar to previously reported Irs1 alleles, mice exhibited decreased survival, in contrast to the enhanced longevity reported for IRS1 deficiency generated by gene targeting. This discrepancy could result from differences in strain background, unintended indirect effects of the gene targeting, or the minimal genetic interference of the S57X mutation compared with the conventionally targeted Irs1-KO allele. Spontaneous mutations arising during ES cell culture may be a frequent but underappreciated occurrence. When linked to a targeted allele, such mutations could lead to incorrect assignment of phenotype and may account for a subset of markedly discordant results from experiments independently targeting the same gene.
机译:在表征具有靶向破坏SerpinB2基因的小鼠的过程中,我们观察到了一些小动物,它们出生时发育迟缓,寿命缩短。尽管此表型与无活性SerpinB2等位基因的纯合性共隔离,但分析来自两个另外的独立胚胎干(ES)细胞克隆的纯合SerpinB2缺陷小鼠的生长无异常。从原始的5erpinB2缺陷品系检查了其他后代,揭示了小表型(smla)与SerpinB2基因座之间的重组。将负责smla的基因座定位在SerpinB2近端大约30 Mb的2.78-Mb区间,以标记D1Mit382和D1Mit216为界。 Irs1的测序确定了在丝氨酸57(SS7X)的无意义突变,导致IRS1蛋白表达的完全丧失。 ES细胞DNA的分析表明,在细胞培养过程中,ES细胞亚克隆中自发出现S57X Irs1突变。尽管smla表型与先前报道的Irs1等位基因相似,但小鼠表现出存活率降低,这与基因靶向产生的IRS1缺乏症报道的寿命延长有关。与常规靶向的Irs1-KO等位基因相比,这种差异可能是由于菌株背景的差异,基因靶向的意外间接作用或S57X突变的最小遗传干扰所致。 ES细胞培养过程中发生的自发突变可能是常见的,但未被充分认识。当与目标等位基因连锁时,此类突变可能导致表型分配不正确,并可能解释了独立针对相同基因的实验产生的明显不一致结果的子集。

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  • 作者单位

    Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109 Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109;

    rnDepartment of Genetics, University of North Carolina, Chapel Hill, NC 27599;

    rnDepartments of Internal Medicine, Human Genetics, and Pediatrics, University of Michigan, Ann Arbor, MI 48109;

    rnHoward Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109;

    rnDepartments of Internal Medicine, Human Genetics, and Pediatrics, University of Michigan, Ann Arbor, MI 48109;

    rnHoward Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109;

    rnDepartments of Internal Medicine, Human Genetics, and Pediatrics, University of Michigan, Ann Arbor, MI 48109;

    rnDepartments of Internal Medicine, Human Genetics, and Pediatrics, University of Michigan, Ann Arbor, MI 48109;

    rnLife Sciences Institute, University of Michigan, Ann Arbor, MI 48109 Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109 Departments of Internal Medicine, Human Genetics, and Pediatrics, University of Michigan, Ann Arbor, MI 48109;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    insulin receptor substrate protein; mutant strain; plasminogen activator inhibitor 2; knockout mice; embryonic stem cell mutation;

    机译:胰岛素受体底物蛋白;突变株纤溶酶原激活物抑制剂2;剔除小鼠胚胎干细胞突变;

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