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）：众所周知，一定比例的转基因插入导致与实际转基因无关的可识别表型。 这是基因组重排的结果，通常是伴随转基因插入的相对较小的缺失和/或反转的结果。 该应用旨在利用这一发现，并系统地识别和表征在影响性别确定和/或生育能力的基因中携带插入突变的小鼠。 研究人员将通过引入基因组中的酪氨酸酶微酸酶的引入酪氨酸酶的近交FVB/N菌株中的白化病来使用新型的转基因插入诱变方案。 根据他们的经验，使用这种方法从小规模的诱变程序中获得，10-15％的插入导致了新的可识别突变，其中10-15％涉及生殖适应性，包括精子发生，卵子发生，性别确定和分娩。 这种方法比使用化学诱变来鉴定生殖突变体的化学诱变具有许多优势。 整个筛选过程，包括将纯合子与杂合子区分开来，可以通过简单的外套颜色进行视觉检查来实现。 它在整个过程中都使用单个近交菌株，消除了任何混杂的遗传背景影响。 与ENU诱变不同，它不需要使用复杂的局部反转或平衡染色体来帮助维持无菌突变体。 最后，其最吸引人的特征之一是该突变被转基因标记。 这允许对破坏基因座的细胞遗传学映射和快速分子分析。 鉴于该新计划在产生生殖突变体以及公共领域中小鼠基因组序列的可用性方面的生产性质，研究人员建议以系统的方式检查更多的转基因小鼠。 不仅将详细识别并表现出生殖突变体，而且将获得其细胞遗传学图位置，产生侧翼序列，并确定突变的确切位置。 博士。 Colin Bishop和Paul Overbeek过去在分析具有多种生殖表型的转基因插入突变体方面经验丰富。 与女性生殖生物学领域的公认专家多洛雷斯·兰姆（Dolores Lamb）以及女性繁殖领域的专家马丁·马祖克（Martin Matzuk）博士一起，研究人员认为拟议的计划将非常有生产力。 该计划设计为国家资源，应为生殖遗传学领域的研究人员提供独特且高度详细的数据库，并访问新的小鼠菌株。 使用不同的策略，它也将与其他诱变计划相辅相成。