Tol2-mediated gene and enhancer trapping in Xenopus tropicalis

热带爪蟾中 Tol2 介导的基因和增强子捕获

• 批准号: 7558192
• 负责人: PAUL E MEAD
• 金额: $ 41.47万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-05 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7558192
• 关键词: Adult; Alleles; Animals; Automobile Driving; Breeding; Cells; Chimeric Proteins; Cloning; Data; Databases; Embryo; Enhancers; Enzymes; Event; Eye; Family; Female; Figs - dietary; Frequencies; Future; Generations; Genes; Germ Lines; Gifts; Hand; Heat-Shock Response; Individual; Injection of therapeutic agent; Japan; Keratin; Manuscripts; Mediating; Messenger RNA; Molecular Profiling; Online Systems; Pattern; Polyadenylation; Protocols documentation; Rana; Records; Relative (related person); Reporter; Seeds; Site; Staging; System; Tadpoles; Time; Tissues; Transgenes; Transgenic Organisms; Transposase; Work; Xenopus; Zebrafish; base; egg; interest; male; novel; promoter; protamine 1; research study; scaffold; sperm cell; transmission process; vector; zygote

We have made important progress on the Tol2 project since the revised application and the supplemental data were submitted in 2008. The most significant advance is the demonstration of germ line transmission of remobilized Tol2 transposons following the injection of mRNA encoding Tol2 transposase in fertilized eggs insertion event (Fig. 1). We had previously shown that somatic remobilization of Tol2 transposons is very efficient; 100% of injected embryos have novel reintegration events following injection of Tol2 transposase mRNA. Here, we demonstrate that the remobilization events are passed through the germ line and can produce tadpoles with novel GFP expression profiles. Figure 1 depicts the analysis of a single remobilized F2 animal (12M2r􀀁5). Three of the four remobilization events found in 12M2r􀀁5 have integrated into interesting gene loci (jvh, cen, fol). Outcross of other F2 remobilized frogs has indicated germ line transmission of novel re-integration events highlighted by the non-Mendelian frequencies of GFP-positive tadpoles in the F3 generation; that is, we observed a significant deviation from the expected 50% GFP-positive progeny from the outcross of remobilized frogs heterozygous for the Tol2XIG transposon. We are currently characterizing the novel integration events in the progeny from the other remobilized F2 frogs. We anticipate submitting a manuscript describing this data soon.

自2008年提交修订申请和补充数据以来，我们在港岛二号工程方面取得了重要进展。最重要的进展是证明了重新激活的Tol2转座子的种系传播