Sleeping Beauty for mouse genetic research

描述（由申请方提供）：睡美人（SB）转座因子将用于小鼠的染色体工程、区域诱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位，这将有助于遗传筛选，突变克隆和菌株维护。去除肿瘤抑制因子的缺陷的产生也将有助于人类肿瘤的建模。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使用Skames实验室开创的含LoxP的基因陷阱进行SB介导的诱变。区域诱变将有助于疾病基因的鉴定，并建立人类疾病的小鼠模型。第三，SB将用于鉴定在癌症起始和进展中与肿瘤抑制因子p19 Arf（Aft）的缺失协作的基因。这将通过在Arf -/-小鼠中动员致突变转座子来实现。将鉴定在比Arf-/-小鼠更早的年龄发生肿瘤的具有新转座子插入的Arf-/-小鼠。的位置 将通过PCR确定肿瘤中的SB插入，并鉴定受影响的基因。总之，这些项目将探索使用SB作为小鼠遗传研究的工具，作为染色体工程，种系诱变和体细胞诱变的手段。

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.