L1 mutagenesis for mammalian models of human diseases

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反转录转座子进行直接种系诱变。转座原的目标是制造“突变鼠”。在这些大鼠中，精子会发生突变，一些后代会含有单基因破坏。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。我们已经开发了“突变小鼠”，其中含有能够种系插入突变的标记L1转基因。本提案的重点是验证转座原随机突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特征。具体目的1是确定突变小鼠精子中插入突变的频率。具体目标是确定插入基因的百分比。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，人类L1元件能够在人类、小鼠、仓鼠、猫甚至鹌鹑细胞中进行高水平的逆转录。因此，我们在小鼠身上的研究结果可以直接外推到大鼠身上。转座原的随机诱变系统最终可能适用于任何哺乳动物。突变小鼠和大鼠将通过产生新的药物靶点的动物模型，极大地帮助药物发现计划。

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.