TRANSGENIC MUTANTS AFFECTING SEX DETERMINATION AND FERT*

影响性别决定和 FERT 的转基因突变体*

• 批准号: 7105576
• 负责人: COLIN Edward BISHOP
• 金额: $ 7.19万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-27 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7105576
• 关键词: albinism; animal colony; biotechnology; cooperative study; cytogenetics; fertility; gene mutation; genetic mapping; genetic strain; genetically modified animals; laboratory mouse; mutant; sex determination

DESCRIPTION (provided by applicant): It is known that a percentage of transgenic insertions result in an identifiable phenotype unrelated to the actual transgene used. This is the result of a genomic rearrangement, usually a relatively small deletion and/or inversion, that accompanies the transgene insertion. This application is designed to exploit this finding and systematically identify and characterize mice carrying insertional mutations in genes affecting sex determination and/or fertility. The investigators will use a novel transgenic insertional mutagenesis scheme based on the rescue of albinism in the inbred FVB/N strain by the introduction a tyrosinase minigene into the genome. In their experience, gained from a small scale mutagenesis program using this method, 10-15% of insertions resulted in new identifiable mutations, of which 10-15% involve reproductive fitness, including spermatogenesis, oogenesis, sex determination, and parturition. This approach offers many advantages over phenotype-driven schemes employing chemical mutagenesis for the identification of reproductive mutants. The entire screening process, including distinguishing homozygotes from heterozygotes, can be achieved by simple visual inspection of coat color. It uses a single inbred strain throughout, eliminating any confounding genetic background effects. Unlike ENU mutagenesis, it does not require the use of complicated local inversions or balancer chromosomes to help maintain sterile mutants. Finally, one of its most attractive features is that the mutation is tagged by the transgene. This allows cytogenetic mapping and fast molecular analysis of the disrupted locus. Given the previous productive nature of this novel program with respect to producing reproductive mutants, and the availability of the mouse genome sequence in the public domain, the investigators propose to examine a greater number of transgenic mice in a systematic way. Not only will reproductive mutants be identified and phenotyped in detail, but their cytogenetic map position will be obtained, flanking sequences generated, and the exact position of the mutation in the mouse genome determined. Drs. Colin Bishop and Paul Overbeek have had considerable experience in the past in analyzing transgenic insertion mutants exhibiting a variety of reproductive phenotypes. Working together with Dr. Dolores Lamb, an acknowledged expert in the field of male reproductive biology, and Dr. Martin Matzuk, an expert in the field of female reproduction, the investigators feel that that the proposed program will be very productive. Designed as a national resource, this program should provide researchers in the field of reproductive genetics a unique and highly detailed database and access to novel strains of mice. It will also be highly complementary to other mutagenesis programs using different strategies.

描述（由申请人提供）：已知一定百分比的转基因插入导致与实际使用的转基因无关的可识别表型。 这是伴随转基因插入的基因组重排的结果，通常是相对小的缺失和/或倒位。 本申请旨在利用这一发现，并系统地鉴定和表征在影响性别决定和/或生育力的基因中携带插入突变的小鼠。 研究人员将使用一种新的转基因插入诱变方案，该方案基于通过将酪氨酸酶小基因引入基因组来拯救近交系FVB/N菌株中的白化病。 根据他们的经验，从使用这种方法的小规模诱变程序中获得的10-15%的插入导致新的可识别的突变，其中10-15%涉及生殖适应性，包括精子发生、卵子发生、性别决定和分娩。 这种方法提供了许多优势，表型驱动的计划，采用化学诱变鉴定生殖突变体。 整个筛选过程，包括区分纯合子和杂合子，可以通过简单的毛色目视检查来实现。 它使用一个单一的近交系在整个，消除任何混淆的遗传背景效应。 与ENU诱变不同，它不需要使用复杂的局部倒位或平衡染色体来帮助维持不育突变体。 最后，它最吸引人的特征之一是突变被转基因标记。 这允许对破坏的基因座进行细胞遗传学作图和快速分子分析。 考虑到这一新计划在生产生殖突变体方面的先前生产性，以及小鼠基因组序列在公共领域的可用性，研究人员建议以系统的方式检查更多数量的转基因小鼠。 不仅将生殖突变体进行详细鉴定和表型分析，而且还将获得它们的细胞遗传学图谱位置，生成侧翼序列，并确定突变在小鼠基因组中的确切位置。 Colin Bishop博士和Paul Overbeek博士过去在分析表现出各种生殖表型的转基因插入突变体方面具有相当丰富的经验。 与男性生殖生物学领域公认的专家Dolores Lamb博士和女性生殖领域的专家Martin Matzuk博士合作，研究人员认为拟议的计划将非常富有成效。 作为一个国家资源，该计划应该为生殖遗传学领域的研究人员提供一个独特的和非常详细的数据库，并获得新的小鼠品系。 它也将与使用不同策略的其他诱变程序高度互补。