SATURATION MUTAGENESIS OF ZEBRAFISH COAGULATION PATHWAY

斑马鱼凝血途径的饱和诱变

• 批准号: 6197922
• 负责人: PUDUR JAGADEESWARAN
• 金额: $ 21.68万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6197922
• 关键词: activation product; blood coagulation; coagulation factor VII; complementary DNA; developmental genetics; fibrinogen; gene expression; gene mutation; genetic models; genome; hemostasis; linkage mapping; molecular biology information system; mutant; phenotype; polymerase chain reaction; thrombin; thromboplastin; thrombosis; zebrafish

DESCRIPTION: (Investigator's abstract) The initiation of blood coagulation through the extrinsic pathway is a threshold-mediated event influenced by relative levels of factor VIIa, tissue factor and TFPI activity. Despite intensive study, important questions remain regarding initiation and regulation of the coagulation response. The applicant hypothesizes that novel genes, involved in initiation and regulation of the extrinsic pathway of coagulation, remain to be identified. The current proposal employs the zebrafish as a genetic tool to identify novel genes in this pathway. The novel genes will be identified by chemical mutagenesis of the zebrafish genome followed by a comprehensive screen for mutants with defective thrombin generation by the extrinsic pathway. Screening for zebrafish phenotypes with defective thrombin generation will yield a significant number of mutations in both known and unknown genes involved in the extrinsic pathway of coagulation. Zebrafish that display defective thrombin generation will be rapidly sorted by both biochemical and linkage analyses to eliminate mutations in known genes, and focus will be placed on novel defects affecting the expression, activation or activity of factor VIIa. Identification of novel zebrafish genes will ultimately allow identification of human homologues involved in hemostasis and thrombosis. Identification of human homologues will further our understanding of how the coagulation cascade functions in vivo, suggest novel targets for antithrombotic therapy and provide candidate genetic markers for thrombotic disease.

描述：（研究者摘要）凝血开始 通过外源性途径是一个阈值介导的事件， 因子VIIa、组织因子和TFPI活性的相对水平。尽管 深入研究，关于启动和调节的重要问题仍然存在 凝血反应。申请人假设新基因， 参与外源性凝血途径的启动和调节， 仍有待确认。目前的提议将斑马鱼作为一种 基因工具，以确定新的基因在这一途径。新的基因将是 通过对斑马鱼基因组进行化学诱变， 通过凝血酶生成缺陷的突变体的综合筛选， 外在途径斑马鱼凝血酶缺陷型的筛选 这一代人将在已知和未知的基因中产生大量的突变， 参与凝血外源性途径的未知基因。斑马鱼 显示有缺陷的凝血酶产生将被快速分类， 生物化学和连锁分析，以消除已知基因中的突变， 重点将放在影响表达，激活或 因子VIIa的活性。新的斑马鱼基因的鉴定将 最终允许鉴定参与止血的人类同源物， 血栓形成人类同源物的鉴定将进一步加深我们对 如何凝血级联功能在体内，提出了新的目标， 抗血栓治疗，并提供血栓形成的候选遗传标记物 疾病