Genetic and genomic tools for C. briggsae research

遗传和基因组工具用于桥冠藻研究：项目摘要 利用微小线虫线虫的研究已经产生了许多基础知识 在细胞死亡的遗传基础、组织衰老、microRNA的生物学作用以及 与人类健康相关的其他基本话题。这是对C. 线虫是那些与线虫有亲缘关系的线虫，它分享了许多实验上的 线虫的优势，但它在大约3000万年前背离了线虫。C.briggsae提供了一个 比较遗传学研究平台，导致对保守过程的有效分析，以及 关于基因、途径和网络进化的发现。这些比较研究在 建立严谨的研究并验证研究结果。此外，线虫和线虫之间的差异也不明显。 生活史特征和全球人口分布方面的重大变化意味着许多研究都在解决 群体生物学问题、多基因特征或宿主-病原体、共生关系和机会性关系 与病毒、细菌和昆虫接触时，与线虫相比，更好地处理布里奇萨线虫。尽管如此 重要特征，许多对标准线虫研究必不可少的遗传资源不是 可用于使用C.briggsae的研究。随着基因编辑和工程应用的效率不断提高 对于CRISPR介导的方法，重要的基因组和遗传工具的可用性是这些方法的关键限制 重要的比较研究。 该项目将产生这些研究资源来解决这一差距。作为第一个目标，两个端粒对- 将使用几套端粒C.briggsae参考基因组和验证的基因模型来生产 互补的长序列和短序列读取方法和验证技术。这些数据将是 纳入NCBI和Wormbase，这是研究人员使用线虫和线虫的在线数据库 相关线虫。第二个目标是生产和验证一组基因平衡器，或重新排列 防止减数分裂交叉的染色体。这些遗传工具对于维护和 对纯合、致命或不育的突变进行评估，这些突变目前通过 费力的过程。基因平衡器还将允许目前还没有的更复杂的基因实验 在C.briggsae是可行的。最后，将具有“避风港”登陆地点的菌株导入明确的DNA 将产生和验证布里奇萨弧菌基因组中的位置。这些菌株将允许受控， 可重复性地将单拷贝插入克隆引入到C.briggsae中，并允许一系列实验 操纵，包括两个物种之间的基因“节点交换”，以及为 秀丽隐杆线虫直接在布氏隐杆线虫中传播。这些目标加在一起，将产生消除重要障碍的关键工具 在这个研究有机体中进行遗传分析。

Genetic and Genomic tools for C. briggsae research: Project Summary Research using the microscopic nematode Caenorhabditis elegans has produced many foundational discoveries in the genetic basis of cell death, organismal aging, the biological roles for microRNAs, as well as other fundamental topics that are relevant to human health. An important complement to studies on C. elegans is those on the related nematode Caenorhabditis briggsae, which shares many of the experimental strengths of C. elegans, but from which it diverged approximately 30 million years ago. C. briggsae provides a platform for comparative genetic studies, leading to efficient analysis of conserved processes, as well as discoveries on the evolution of genes, pathways and networks. These comparative studies are important in establishing research rigor and validating findings. In addition, differences between C. elegans and C. briggsae in terms of life history traits and global distribution of populations means that many studies addressing population biology questions, polygenic traits, or host-pathogen, commensal, and opportunistic relationships with viruses, bacteria, and insects are better done with C. briggsae than with C. elegans. Despite these important features, many genetic resources that are essential for standard C. elegans research are not available for studies using C. briggsae. With the increasing efficiency of genetic editing and engineering using CRISPR-mediated methods, the availability of important genomic and genetic tools is a key limitation for these important comparative studies. This project will produce these research resources to address this gap. As a first aim, two telomere-to- telomere C. briggsae reference genomes with validated gene models will be produced, using sets of complementary long and short sequence read methods and validation techniques. These data will be incorporated into NCBI and Wormbase, the online database used by researchers who use C. elegans and related nematodes. A second aim will produce and validate a set of genetic balancers, or rearranged chromosomes that prevent meiotic crossing over. These genetic tools are critical for the maintenance and evaluation of mutations that are homozygous lethal or sterile and that are currently maintained through laborious processes. Genetic balancers will also permit more complex genetic experiments not currently feasible in C. briggsae. Finally, strains with "safe harbor" landing sites for the introduction of DNA into defined locations in the C. briggsae genome will be produced and validated. These strains will permit controlled, reproducible introduction of single copy insertion clones into C. briggsae, and permit a range of experimental manipulation including gene "node swaps" between the two species, and the testing of reagents developed for C. elegans directly in C. briggsae. Together, these aims will produce key tools that remove important barriers to genetic analysis in this research organism.