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 逆转录转座子进行直接种系诱变。 Transposagen 的目标是培育“突变老鼠”。在这些大鼠中，精子中会发生诱变，并且一些后代将含有单基因破坏。我们将使用小鼠来优化和验证我们的系统，因为生产转基因大鼠的成本要高得多。我们已经开发出含有能够进行种系插入诱变的标记 L1 转基因的“突变小鼠”。该提案的重点是验证 Transposagen 随机诱变系统作为功能基因组学工具的作用。该项目的第一阶段包括突变小鼠的表征。具体目标1是确定突变小鼠精子中插入突变的频率。具体目标 2 是确定插入基因的插入百分比。具体目标 3 是确定基因破坏的效率。该项目的第二阶段将利用从第一阶段小鼠研究中获得的信息来创建突变大鼠。 L1 元件存在于所有哺乳动物中，人类 L1 元件能够在人类、小鼠、仓鼠、猫科动物甚至鹌鹑细胞中进行高水平逆转座。因此，我们在小鼠身上的研究结果应该可以直接推断到大鼠身上。 Transposagen 的随机诱变系统最终可以适用于任何哺乳动物。突变小鼠和大鼠将通过制作新药物靶点的动物模型来极大地帮助药物发现计划。