Sleeping Beauty for mouse genetic research

描述(由申请人提供)：睡美人(SB)转座元件将用于小鼠的染色体工程、区域突变和癌症遗传学研究。首先，我们将使用噬菌体C31整合酶、SB和Cre-loxP在小鼠中进行染色体工程。我们将创造染色体缺陷和倒置，这将有助于基因筛查、突变克隆和菌株维护。去除肿瘤抑制基因的缺陷的产生也将有助于对人类肿瘤进行建模。其次，Sb将用于局部诱变。与人类疾病区域同源的小鼠染色体的特定区域将被定位于SB介导的突变，使用斯凯姆斯实验室首创的含有loxP的基因陷阱。区域突变将促进疾病基因的识别，并创建人类疾病的小鼠模型。第三，SB将被用来识别与肿瘤抑制基因p19Arf(Aft)丢失在癌症发生和发展中协同作用的基因。这将通过在Arf-/-小鼠中动员诱变转座子来实现。具有新的转座子插入的ARF-/-小鼠将被识别出来，这些小鼠比ARF-/-小鼠更早患上肿瘤。的位置 肿瘤中的SB插入将通过聚合酶链式反应来确定，并鉴定受影响的基因。总之，这些项目将探索将SB作为小鼠遗传学研究的工具，作为染色体工程、生殖系突变和体细胞突变的手段。

DESCRIPTION (provided by applicant): The Sleeping Beauty (SB) transposable element will be used for chromosome engineering, regional mutagenesis, and cancer genetic studies in the mouse. First, we wilt use phage C31 integrase, SB, and CRE-LoxP for chromosomal engineering in the mouse. We will create chromosomal deficiencies and inversions that will be useful for genetic screens, cloning of mutations, and strain maintenance. The generation of deficiencies that remove tumor suppressors will also be useful for modeling human tumors. Second, SB will be used for local mutagenesis. Specific regions of mouse chromosomes homologous to human disease regions will be targeted for SB-mediated mutagenesis using LoxP-containing gene traps pioneered by the Skames laboratory. Regional mutagenesis will facilitate identification of disease genes and create mouse models of human disease. Third, SB will be used to identify genes that cooperate with loss of the tumor suppressor p 19Arf (Aft) in cancer initiation and progression. This will be accomplished by mobilizing mutagenic transposons in Arf -/- mice. Arf-/- mice with novel transposon insertions that develop tumors at an earlier age than Arf-/- mice will be identified. The location of SB insertions in the tumors will be determined by PCR, and the affected gene identified. Together, these projects wilt explore the use of SB as a tool for mouse genetic research as a means for chromosome engineering, germline mutagenesis, and somatic mutagenesis.