Embryonic Functional Screen of a TGF?? Protein-Protein Interaction Network

TGF 的胚胎功能筛查？

基本信息

批准号：
8069332
负责人：
GERALD H THOMSEN
金额：
$ 7.54万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2014-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8069332
关键词：
Affect Affinity Biochemical Biological Biological Assay Biological Process Cancer Center Support Grant Candidate Disease Gene Cell Differentiation process Cloning Co-Immunoprecipitations Complementary DNA Complex Congenital Abnormality Data Databases Development Developmental Process Disease Embryo Embryonic Development Essential Genes Expressed Sequence Tags Gene Expression Genes Genetic Translation Genome Genomics Goals Grant Growth Factor Human Injection of therapeutic agent Messenger RNA Neurula Nodal Oligonucleotides Organism Pathway interactions Phenotype Play Process Property Proteins Publishing RNA Splicing Relative (related person)Research Personnel Role Screening procedure Signal Transduction Source Staging System Systems Biology Testing Time Tissues Transforming Growth Factor beta Translations Wound Healing Xenopus Xenopus laevis base egg loss of function mRNA Precursor novel overexpression protein protein interaction public health relevance research study stem cell division time use

项目摘要

DESCRIPTION (Provided by Applicant): The field of Systems Biology is beginning to define the complex dynamics of biological regulatory processes. Recently, large-scale protein-protein interaction (PPI) screens have been performed on the TGF-¿ signaling system, revealing a complicated network of hundreds of interacting proteins, many novel. A key question is whether biological processes governed by TGF-¿ signals are regulated by these newfound proteins. TGF-¿ signals govern many important biological processes, but particularly cell differentiation and embryonic development. Therefore, large-scale PPI screens are potentially a rich source of new developmental regulators, but new candidates must be evaluated in an embryonic context. The investigator proposes to use Xenopus embryos to screen new TGF-¿ signaling candidates for developmental activity. His hypothesis is that embryonic functional assays will uncover novel developmental regulators among new interacting proteins emerging from TGF-¿ PPI screens. The approach is to test these proteins for activity in Xenopus embryos by overexpression and gene knockdown. Aim 1 will evaluate the results of published and ongoing PPI screens on the TGF-¿ system to select novel candidates for embryonic tests. Evidence of candidate expression in Xenopus embryos will be initially assessed by EST database searches, or by cDNA cloning. Sequence information will be gathered for genes expressed at neurula or earlier times using EST databases, cDNA cloning, and Xenopus tropicalis genomic information. Aim 2 will perform candidate gene knockdown in embryos with morpholino oligos that block mRNA translation or pre-mRNA splicing. These tests will reveal new developmental regulators and expand our understanding of TGF-¿ signaling and embryogenesis. In broader context, new regulators to be uncovered will inform various fields of new genes that could affect birth defects and diseases through abnormal expression or function, particularly in TGF-¿ signaling. PUBLIC HEALTH RELEVANCE: Growth factor signals, such as those provided by relatives of TGF-¿, govern many important biological processes, such as cell differentiation, embryonic development, wound healing, and tissue/stem cell renewal. The tests proposed in this application will reveal potential functions of new regulators of TGF-¿ signaling during vertebrate embryogenesis. The experiments will point to new genes that might affect human developmental processes, and which might cause birth defects and disease if their activity or expression is abnormal.

描述（由申请人提供）：系统生物学领域开始定义生物调节过程的复杂动力学。最近，已经在TGF-¿上进行了大规模蛋白质-蛋白质相互作用（PPI）筛选。信号系统，揭示了数百种相互作用蛋白质的复杂网络，其中许多新颖，一个关键问题是生物过程是否受 TGF-¿信号由这些新发现的 TGF-¿信号控制许多重要的生物过程，特别是细胞分化和胚胎发育，因此，大规模 PPI 筛选可能是新发育调节因子的丰富来源，但研究人员建议使用非洲爪蟾胚胎。筛选新的 TGF-¿他的假设是，胚胎功能测定将揭示 TGF-¿ 中出现的新相互作用蛋白中的新发育调节因子。 PPI 筛选。该方法是通过过度表达和基因敲低来测试这些蛋白质在非洲爪蟾胚胎中的活性，目标 1 将评估已发表和正在进行的 TGF-¿系统选择新的候选胚胎测试。非洲爪蟾胚胎中候选表达的证据将通过 EST 数据库搜索进行初步评估，或通过 EST 数据库、cDNA 克隆收集神经胚或更早时期表达的基因的序列信息。 Aim 2 将使用阻断 mRNA 翻译或前 mRNA 剪接的吗啉寡核苷酸在胚胎中进行候选基因敲除。这些测试将揭示新的发育调节因子并进行扩展。我们对 TGF-¿ 的理解在更广泛的背景下，待发现的新调控因子将告知新基因的各个领域，这些基因可能通过异常表达或功能影响出生缺陷和疾病，特别是在 TGF-¿发信号。公共卫生相关性：生长因子信号，例如 TGF-¿ 亲属提供的信号，控制许多重要的生物过程，例如细胞分化、胚胎发育、伤口愈合和组织/干细胞更新。本申请中提出的测试将揭示 TGF-¿ 的新调节剂的潜在功能。这些实验将指出可能影响人类发育过程的新基因，如果它们的活性或表达异常，可能会导致出生缺陷和疾病。