The Evolution of Posterior Fate

体型和体轴的进化依赖于调控网络的变化，这些网络控制着整个生物体的组织发育。这笔赠款将研究一个物种的基因调控网络的变化如何改变，并导致其他物种的形态新颖性的形成。揭示物种之间形态多样性背后的自然变化将有助于阐明器官形成策略。初步结果表明，基因组工程技术在非传统物种中的成功应用，将使首席研究者能够将它们建立为新的模式系统，以研究基因调控的变化如何控制新形态的进化。所研究的信号通路和转录因子在其他动物中是保守的，因此果蝇群落和其他发育群落将获得关于这些通路的知识，并从产生的遗传工具中受益。一名博士后实习生、一名博士生和9名本科生将接受苍蝇遗传学、分子生物学、生物信息学和基因组工程方面的培训，重点是来自STEM领域未被充分代表的少数民族的学员。生成的数据将被纳入研究生和本科生水平的遗传学课程，每年为大约60名学生提供服务。在生物学访问日期间，首席调查员将与来自新泽西州卡姆登地区的未来生物学学生及其家人进行接触。轴的形成是动物发育的基本要求。这一复杂的过程受动物体内少数高度保守的细胞信号通路的调控。首席研究员(PI)将研究由表皮生长因子受体(EGFR)、Janus Kinase(JAK/STAT)、骨形态发生蛋白(BMP)和控制果蝇前后轴的转录调控因子组成的基因调控网络的进化。等电点表明，该网络的组成部分从控制苍蝇卵子发生过程中轴的形成，演变为也调节着云杉卵壳上一种形态上的新颖物--背脊的形成。PI的实验室通过发现不同物种之间EGFR激活模式的戏剧性变化，开创了背脊形成的分子分析。PI假设，在一个物种中，控制后天命运的基因顺式调控的变化是为了指导其他物种中新形态的产生。初步结果表明，Ets转录因子定点(PNT)控制后决定簇中线(Mid)的表达。研究目标是，a)确定控制后端模式的网络，b)研究控制MID表达的顺式调控模块，以及3)研究MID模式的变化是否控制背脊的进化。使用免疫染色、遗传扰动(包括CRISPR/Cas9)和同源CRM的互换的组合，这些CRM的功能、要求和冗余将在黑腹果蝇和星云果蝇中进行图案化和形态发生测试。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

The evolution of body shape and body axes relies on changes to regulatory networks that control tissues' development across organisms. The grant will examine how changes in the regulatory network of genes in one species can be changed and lead to the formation of morphological novelties in other species. Uncovering the natural changes underlying morphological diversity among species will shed light on organ formation strategies. The preliminary results demonstrate the successful use of a genome engineering technique in non-traditional species, which will allow the principal investigator to establish them as new model systems to study how changes in gene regulation control the evolution of new morphologies. The studied signaling pathways and transcription factors are conserved in other animals, hence the Drosophila community and other developmental communities will gain knowledge on these pathways and benefit from the generated genetic tools. A Postdoctoral trainee, a Ph.D. student, and 9 undergraduate students will be trained in fly genetics, molecular biology, bioinformatics and genome engineering, with an emphasis on trainees from underrepresented minorities in the STEM fields. The data generated will be incorporated into the Genetics class curricula at the level of graduate and undergraduate studies, which serves about 60 students annually. The principal investigator will reach out to prospective biology students and their families from the Camden, NJ area during the biology visiting days.Axis formation is a fundamental requirement during animal development. This intricate process is regulated by a small number of highly-conserved cell signaling pathways across animals. The principal investigator (PI) will study the evolution of gene regulatory networks, comprised of the epidermal growth factor receptor (EGFR), the Janus Kinase (JAK/STAT), bone morphogenetic protein (BMP), and transcriptional regulators that control the anterior posterior axis in Drosophila melanogaster. The PI suggests that components of the network evolved from controlling axis formation during fly oogenesis to also regulating the formation of a morphological novelty, the dorsal ridge, on the D. nebulosa eggshells. The PI's laboratory pioneered the molecular analysis of dorsal ridge formation by discovering the dramatic changes in EGFR activation patterns among species. The PI hypothesizes that changes in cis regulation of genes controlling the posterior fate in one species evolved to guide the generation of a new morphology in other species. The preliminary results demonstrate that the ETS-transcription factor Pointed (PNT) controls the expression of the posterior determinant Midline (Mid). Research goals are to, a) determine the network controlling patterning of the posterior end, b) study the cis regulatory modules controlling mid expression and 3), study whether changes in MID patterning control the evolution of the dorsal ridge. Using a combination of immunostaining, genetic perturbations, including CRISPR/Cas9, and reciprocal swapping of orthologous CRMs, the function, requirement, and redundancy of these CRMs will be tested for patterning and morphogenesis in both D. melanogaster and D. nebulosa.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.