A Screen for DNA Damage Checkpoint Mutants in Zebrafish

斑马鱼 DNA 损伤检查点突变体的筛选

• 批准号: 6937498
• 负责人: CHRISTOPHER L SANSAM
• 金额: $ 4.4万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6937498
• 关键词: DNA damage; DNA repair; DNA replication; alleles; animal population genetics; biological signal transduction; cancer risk; cell cycle proteins; cell growth regulation; cell line; embryo /fetus; gene expression; genetic library; genetic screening; genetic susceptibility; human tissue; laboratory mouse; mutant; neoplasm /cancer genetics; neoplastic cell; postdoctoral investigator; protein structure function; tissue /cell culture; zebrafish

DESCRIPTION (provided by applicant): Cancer cells are created through accumulated DNA mutations in somatic cells, where cellular DNA is constantly assaulted by exogenous agents and cellular metabolites. Normal cells have mechanisms for monitoring genomic DNA integrity and repairing the DNA mutations, while cancer cells frequently lose the ability to efficiently respond to DNA damage, facilitating the accumulation of mutations that lead to deregulated growth. A critical cellular response to DNA damage is the arrest of the cell cycle, thereby providing time for repair before DNA replication or mitosis. The mechanisms of DNA damage-induced cell cycle arrest are complex, and many components of the signal transduction pathway are unknown. The zebrafish has considerable advantages as a vertebrate model for the discovery of DNA damage response genes. By screening zebrafish embryos in the Hopkins' insertional mutant collection, two recessive embryonic lethal mutations were identified that abrogate DNA damage-induced cell cycle arrest. The genes affected by these mutations have been cloned, and they have human homologues with high similarity, but little is known about their functions. The function of these genes in the DNA damage checkpoints of human cells will be examined. Also, the Hopkins' mutant collection will be screened for additional DNA damage checkpoint failure mutants.

描述（由申请人提供）：癌细胞是通过体细胞中积累的 DNA 突变产生的，其中细胞 DNA 不断受到外源物质和细胞代谢物的攻击。正常细胞具有监测基因组 DNA 完整性和修复 DNA 突变的机制，而癌细胞经常失去有效应对 DNA 损伤的能力，从而促进突变的积累，从而导致生长失调。细胞对 DNA 损伤的关键反应是细胞周期的停滞，从而在 DNA 复制或有丝分裂之前提供修复时间。 DNA损伤引起细胞周期停滞的机制很复杂，信号转导途径的许多组成部分尚不清楚。斑马鱼作为发现DNA损伤反应基因的脊椎动物模型具有相当大的优势。通过筛选霍普金斯大学插入突变体库中的斑马鱼胚胎，发现了两种隐性胚胎致死突变，它们可以消除 DNA 损伤诱导的细胞周期停滞。受这些突变影响的基因已被克隆，它们与人类同源物具有高度相似性，但对其功能知之甚少。将检查这些基因在人类细胞 DNA 损伤检查点中的功能。此外，霍普金斯大学的突变体收藏品还将筛选额外的 DNA 损伤检查点失败突变体。