L1 mutagenesis for mammalian models of human diseases

描述（由申请人提供）：转座蛋白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反转录转座子进行直接生殖系诱变。转座蛋白的目标是生产“突变鼠”。在这些大鼠中，突变将发生在精子中，一些后代将包含单基因破坏。由于生产转基因大鼠的成本要高得多，我们将使用小鼠来优化和验证我们的系统。我们已经开发了“突变小鼠”含有一个显着的L1转基因能够生殖系插入诱变。该提案的重点是验证转座基因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使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，并且人L1元件能够在人、小鼠、仓鼠、猫甚至鹌鹑细胞中进行高水平的反转录转座。因此，我们在小鼠中的研究结果应该直接外推到大鼠。转座蛋白的随机诱变系统可适用于任何哺乳动物。突变小鼠和大鼠将通过产生新药物靶点的动物模型来大大帮助药物发现计划。

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.