Resource for marking clones on the fly 4th chromosome

用于标记飞行中克隆的资源第四染色体 世纪以来，利用黑腹果蝇的研究在我们的研究中取得了重大进展。 理解高度保守的细胞过程和信号系统。在人类健康领域， 果蝇遗传学已成为鉴定疾病相关基因和提供 深入了解其作用机制。第四条染色体（IV）是遗传分析的最后前沿， 果蝇小而缺乏重组的IV在很大程度上被忽视了。然而，第四章的长臂 大约有105个基因这些基因中有55%具有明显的人类直系同源物，67%的人类基因 有一种疾病的关联。要全面了解多细胞性，需要对 这些基因的突变。体细胞克隆和生殖系克隆是用于研究 果蝇的致命突变用于跟踪突变细胞克隆的MARCM系统已被广泛采用 研究中枢神经系统发育和成体肠道干细胞。然而，MARCM是不可用的 由于缺乏合适的染色体，作为果蝇社区的资源， 研究人员几乎由所有的NIH研究所和中心资助，我们提出了一个合作的R21（在 对PAR-16-141的反应）以产生MARCM-IV所需的染色体。我们的两个实验室目前 NIH资助了TGF-β信号传导的研究，并在之前就Sno的作用进行了合作。 癌基因在TGF-β信号转导中的作用我们将采用一种创新的策略， （Crisper-Cas9）与遗传学（X到常染色体跳跃）。然后将在TGF-β研究中测试MARCM-IV 信号在蘑菇体和幼虫大脑的肠神经元。本项目的具体目标 目标1：资源开发：创造MARCM-IV所需的独特的第四染色体。AIM2 发现：主要研究的证据将利用MARCM-IV生成三个突变基因的标记克隆。 它们是TGF-β配体激活素-β和肌球蛋白以及Smad相互作用的信号转导子dCORL （Flybase中的fussel）。MARCM-IV的这些应用将推进我们对分子机制的认识， TGF-β通路，神经胶质细胞相互作用和食管下神经元的发育/功能 个脑袋这一结果应该会引起果蝇群体中其他人的注意， 包含IV上的基因。我们将向任何合格的研究者提供MARCM-IV行，并提供以下证据： 随附于本建议书的信件，已反映了社会人士的关注。鉴于IV上的许多基因 是保守的，新的见解，从MARCM-IV在苍蝇可以很容易地转化为新的假设，正常的 人类的发育/生理或疾病。展望未来，我们的创新方法很可能 扩展到对IV上所有基因的分析。这个项目创造的宝贵社区资源将是 免费提供给所有合格的研究人员，以促进我们了解的保守特征， 发育信号和神经生物学影响人类健康和疾病。

Resource for marking clones on the fly 4th chromosome For over a century, studies employing Drosophila melanogaster have resulted in significant advances in our understanding of highly conserved cellular processes and signaling systems. In the area of human health, Drosophila genetics has been an effective means for identifying disease-associated genes and for providing insights into their mechanism of action. The fourth chromosome (IV) is the final frontier for genetic analysis in Drosophila. Small and devoid of recombination IV has been largely ignored. Nevertheless, the long arm of IV contains roughly 105 genes. 55% of these genes have obvious human orthologs and 67% of the human genes have a disease association. A complete understanding of multicellularity requires the genetic analysis of mutations in these genes. Somatic and germline clones are established tools for studying the functions of lethal mutations in flies. The MARCM system for tracking clones of mutant cells he has been widely employed to study central nervous system development and adult intestinal stem cells. However, MARCM is not useable on IV due to the lack of appropriate chromosomes. As a resource for the Drosophila community whose investigators are funded by virtually all of the NIH Institutes and Centers, we propose a collaborative R21 (in response to PAR-16-141) to generate the necessary chromosomes for MARCM-IV. Our two labs are currently NIH funded for studies of TGF-β signaling and have worked together previously on the role of the Sno oncogene in TGF-β signal transduction. We will employ an innovative strategy integrating molecular methods (Crisper-Cas9) with genetics (X to autosome jumping). MARCM-IV will then be tested in studies of TGF-β signaling in the mushroom body and the enteric neurons of the larval brain. The Specific Aims of this project are: Aim1 Resource Development: To create the unique fourth chromosomes necessary for MARCM-IV. Aim2 Discovery: Proof of principal studies will utilize MARCM-IV to generate marked clones for three mutant genes. These are the TGF-β ligands activin-β and myoglianin and the Smad-interacting signal transducer dCORL (fussel in Flybase). These applications of MARCM-IV will advance our knowledge of molecular mechanisms in the TGF-β pathway, neural-glial interactions and the development/function of subesophageal neurons in the brain. The results should attract the attention of others in the Drosophila community whose interests encompass genes on IV. We will provide the MARCM-IV lines to any qualified investigator and evidence of community interest is already visible in letters appended to this proposal. Given that many of the genes on IV are conserved, new insights from MARCM-IV in flies can be readily translated into new hypotheses for normal development/physiology or diseases in humans. Looking ahead, it is likely our innovative approach can easily be expanded to an analysis of all genes on IV. The valuable community resource created by this project will be made freely available to all qualified researchers to facilitate our understanding of conserved features of developmental signaling and neurobiology impacting human health and disease.