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: 7066040
• 负责人: CHRISTOPHER L SANSAM
• 金额: $ 4.88万
• 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/7066040
• 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）：癌细胞是通过在体细胞中积累的DNA突变创建的，在体细胞中，细胞DNA不断受到外源性剂和细胞代谢产物的攻击。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响应DNA损伤的能力，从而促进了突变的积累，从而导致导致消失受管化的生长。对DNA损伤的关键细胞反应是细胞周期的停滞，从而为DNA复制或有丝分裂提供了修复的时间。 DNA损伤引起的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。通过筛选霍普金斯插入突变体收集中的斑马鱼胚胎，发现两个隐性胚胎致死突变被鉴定出消除DNA损伤引起的细胞周期停滞。受这些突变影响的基因已被克隆，并且具有高相似性的人类同源物，但对其功能知之甚少。将检查这些基因在人类细胞的DNA损伤检查点中的功能。同样，将筛选霍普金斯突变体的收集，以获取额外的DNA损伤检查点故障突变体。