权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The Retinoblastoma corepressor family: precision targeting for discerning gene regulatory mechanisms in the Drosophila eye

视网膜母细胞瘤辅阻遏物家族：精确靶向果蝇眼中辨别基因调控机制

基本信息

批准号：
10456089
负责人：
Ana-Maria Raicu
金额：
$ 3.38万
依托单位：
MICHIGAN STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-16 至 2023-08-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456089
关键词：
Affect Biochemical Pathway Biological Biological Models Chimera organism Chromatin Dependence Development Disease Dissection Drosophila eye Drosophila genome Drosophila genus Embryo Environment Eukaryota Event Exhibits Eye Family Family member Gene Duplication Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Transcription Genome Guide RNA Histones Human Lesion Loss of Heterozygosity Malignant Childhood Neoplasm Malignant Neoplasms Malignant neoplasm of lung Measures Mediating Methods Michigan Modeling Molecular Mus Mutation Normal tissue morphology Pediatric Retinoblastoma Phenotype Phosphorylation Play Post-Translational Regulation Process Property Protein Family Proteins Regulation Regulator Genes Repression Research Retinoblastoma Retinoblastoma Genes Retinoblastoma Protein Role Scientist Site System Technology Testing Tissues Training Transcription Repressor Transcriptional Regulation Tumor Suppressor Genes Tumor Suppressor Proteins Universities Work cancer type career cell type experience fly gene repression gene therapy genetic corepressor improved in vivo malignant breast neoplasm malignant retina neoplasm mutant novel nuclease paralogous gene promoter recruit retinoblastoma tumor suppressor therapeutic development tool transcription factor tumor

项目摘要

PROJECT SUMMARY/ABSTRACT Retinoblastoma is a rare pediatric cancer that is caused by loss of both copies of the retinoblastoma (Rb) tumor suppressor gene. Rb is a highly conserved and critical transcription factor for the proper regulation of gene expression in eukaryotes. Mutations in Rb have also been observed in many human cancers including breast and lung cancer. In humans, the Rb family comprises Rb, p107, and p130, which exhibit both overlapping and non-redundant roles in gene regulation. Similarly, the Drosophila lineage experienced a gene duplication event leading to the expression of the Rbf1 and Rbf2 paralogs. The significance of the multiplicity of the Rb family and their division of labor in regulating gene expression in different contexts is not clear. Their activity has been studied in the context of specific promoters; however, we lack deeper mechanistic understanding of these factors in the context of normal tissues, in development. This proposal will investigate the tissue-specific gene regulatory activities of retinoblastoma family proteins using the fly model system. We hypothesize that the Rb family has diversified to have gene-specific activities through differential targeting to promoters and through different repression activities. In Aim 1, we will investigate how Rb paralogs exhibit mechanistically different transcriptional control while in Aim 2 we will determine how regulation of the Rbf1 protein itself controls promoter-specific repression. We will use our newly developed method for highly precise targeting of Rb paralogs to diverse gene promoters through Rb fusions to a nuclease dead Cas9. This powerful tool allows for direct comparison of the Rb family members in a developmental system to study their gene- and tissue-specific activities. Using this tool, we will determine the differences between Rbf1 and Rbf2 impact on gene expression, and differences in modulation of the chromatin environment. This project will be carried out at Michigan State University in the lab of Dr. David Arnosti as part of a comprehensive academic and professional training plan to prepare the applicant for a career as an independent research scientist investigating gene regulation in the context of disease. With the completion of the proposed work, the results will uncover the mechanisms of gene regulation by Rb family proteins, which will improve our understanding of why certain tissues are especially sensitive to inactivation of Rb. Understanding the mechanisms of Rb-mediated gene regulation and its activity as a transcriptional repressor will allow for the development of gene therapies to treat Retinoblastoma and other cancer types.

项目总结/摘要视网膜母细胞瘤是一种罕见的儿科癌症，是由视网膜母细胞瘤（Rb）肿瘤的两个副本丢失引起的抑制基因Rb是一种高度保守的转录因子，在基因的正确调控中起着关键作用。在真核生物中表达。Rb基因突变在许多人类癌症中也被观察到，包括乳腺癌。和肺癌。在人类中，Rb家族包括Rb、p107和p130，它们表现出重叠和不重叠。在基因调控中的非冗余作用。同样，果蝇谱系也经历了基因复制事件导致Rbf 1和Rbf 2旁系同源物的表达。Rb家族多样性的意义和它们在不同情况下调节基因表达的分工尚不清楚。他们的活动在特定启动子的背景下进行研究;然而，我们缺乏对这些因素更深层次的机制理解在正常组织中，在发育过程中。该提案将研究组织特异性基因调控视网膜母细胞瘤家族蛋白的活性。我们假设Rb家族通过对启动子的差异靶向和通过不同的靶向，镇压活动。在目的1中，我们将研究Rb旁系同源物如何表现出机制上不同的转录水平，而在目标2中，我们将确定Rbf 1蛋白本身的调节如何控制启动子特异性镇压我们将使用我们新开发的方法，高度精确地将Rb旁系同源物靶向到不同的基因，启动子通过Rb融合至核酸酶死亡的Cas9。这个强大的工具允许直接比较 Rb家族成员在发育系统中的作用，以研究其基因和组织特异性活动。使用这个工具，我们将确定Rbf 1和Rbf 2对基因表达的影响之间的差异，以及Rbf 1和Rbf 2对基因表达的影响之间的差异。染色质环境的调节。该项目将在密歇根州立大学的实验室进行大卫Arnosti博士作为一个全面的学术和专业培训计划的一部分，以准备申请人作为一名独立的研究科学家，研究疾病背景下的基因调控。与该工作的完成，将为进一步揭示Rb家族的基因调控机制奠定基础蛋白质，这将提高我们的理解，为什么某些组织是特别敏感的失活， RB.了解Rb介导的基因调控机制及其作为转录抑制因子的活性将允许开发基因疗法来治疗视网膜母细胞瘤和其他癌症类型。