L1 mutagenesis for mammalian models of human diseases

人类疾病哺乳动物模型的 L1 诱变

• 批准号: 6883346
• 负责人: ERIC M OSTERTAG
• 金额: $ 9.97万
• 依托单位: TRANSPOSAGEN BIOPHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-14 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6883346
• 关键词: animal breeding; biomedical resource; biotechnology; cryopreservation; disease /disorder model; functional /structural genomics; gene frequency; gene mutation; gene targeting; genetic manipulation; genetic mapping; genetic transcription; genetically modified animals; laboratory mouse; model design /development; polymerase chain reaction; sperm; tissue resource /registry; transfection; transposon /insertion element

DESCRIPTION (provided by applicant): Transposagen is developing an innovative mammalian random mutagenesis system that uses an L1 retrotransposon for direct germ-line mutagenesis. Transposagen's goal is to produce "mutator rats". In these rats, mutagensis will occur in sperm and some offspring will contain single gene disruptions. We will use the mouse to optimize and validate our system because of the much higher cost in producing transgenic rats. We have developed "mutator mice" containing a marked L1 transgene capable of germ-line insertional mutagenesis. The focus of this proposal is to validate the Transposagen random mutagenesis system as a tool for functional genomics. Phase I of this project includes characterization of the mutator mice. Specific aim 1 is to determine the frequency of insertional mutagenesis in the sperm of mutator mice. Specific aim 2 is to determine the percentage of insertions that insert into genes. Specific aim 3 is to determine the efficiency of gene disruption. Phase II of the project will use the information gained from the Phase I mouse studies to create mutator rats. L1 elements are present in all mammals and a human L1 element is capable of high-level retrotransposition in human, mouse, hamster, feline, and even quail cells. Therefore, our findings in mice from this study should extrapolate directly to rats. Transposagen's random mutagenesis system could utlimately be adapted for use in any mammal. Mutator mice and rats will substantially aid drug discovery programs by producing animal models for new drug targets.

描述（由申请人提供）：Transposagen正在开发一种创新的哺乳动物随机诱变系统，该系统使用L1逆转录座子进行直接种系诱变。 Transposagen的目标是产生“突变器大鼠”。在这些大鼠中，诱变将发生在精子中，一些后代将包含单个基因破坏。我们将使用鼠标来优化和验证我们的系统，因为产生转基因大鼠的成本要高得多。我们开发了包含具有种系插入诱变的明显L1转基因的“突变小鼠”。该提案的重点是验证转泊剂随机诱变系统作为功能基因组学的工具。该项目的第一阶段包括突变器小鼠的表征。具体目的1是确定突变体小鼠精子中插入诱变的频率。具体目标2是确定插入基因的插入的百分比。具体目的3是确定基因破坏的效率。该项目的第二阶段将使用从I期小鼠研究中获得的信息来创建突变器大鼠。 L1元素都存在于所有哺乳动物中，并且人类L1元素能够在人，小鼠，仓鼠，猫甚至鹌鹑细胞中进行高级逆转录。因此，我们在这项研究中对小鼠的发现应直接推断为大鼠。可以将转泊剂的随机诱变系统改编成任何哺乳动物。突变器小鼠和大鼠将通过为新药靶标生产动物模型来实质性地帮助药物发现计划。