FORWARD AND REVERSE SCREENS FOR TEMPERATURE-SENSITIVE MUTATIONS IN ZEBRAFISH

描述（由申请人提供）：尽管斑马鱼领域的快速增长，一个重要的差距是能够容易地评估基因功能超出了第一个合子的基因的要求。条件突变，如温度敏感性突变，在高温或限制性温度（例如，33度）下表达表型，但在低温或允许温度（24度）下正常，允许基因功能在发育的不同时期被剥夺或恢复，以探索基因功能在动物生命周期中不同过程中的贡献。斑马鱼是采用温度敏感策略的理想选择，因为它一生中在24至33度的温度下生长健壮。在这个项目中，我们将使用正向和反向遗传筛选技术来评估识别斑马鱼温度敏感性突变的策略，组装一组100个温度敏感性突变，并采用二次筛选，包括一个由母体效应引起的表型和第二个维持成体干细胞的晚期幼虫缺陷。

DESCRIPTION (provided by applicant): Despite the rapid growth of the zebrafish field, an important gap is the ability to easily assess gene function beyond the first zygotic requirement for the gene. Conditional mutations, such as temperature-sensitive mutations that express phenotypes at a high or restrictive temperature (for instance, 33 degrees) but are normal at a low or permissive temperature (24 degrees) allow gene function to be taken away or restored at different times of development, to explore the contribution of gene function in different processes across the animal's life cycle. The zebrafish is ideal for employing temperature-sensitive strategies, because it grows robustly at temperatures from 24 to 33 degrees throughout its life. In this project, we will use forward and reverse genetic screening techniques to evaluate strategies for identifying temperature-sensitive mutations in zebrafish, assemble a panel of 100 temperature-sensitive mutations, and employ secondary screens, including one for phenotypes caused by maternal effects and a second for late larval defects in maintaining adult stem cells.