Transgenic

转基因

• 批准号: 7945054
• 负责人: Jan M. van Deursen
• 金额: $ 14.21万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-17 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7945054
• 关键词: Alleles; Animal Model; Animals; Applications Grants; Arizona; Autologous; Biological Assay; Biopsy; Breeding; Cell Culture Techniques; Cell Line; Cell Separation; Cell division; Cells; Characteristics; Chimera organism; Chromosomes; Clinic; Cloning Vectors; Collection; Complementary DNA; Comprehensive Cancer Center; Consultations; DNA; DNA Sequence Rearrangement; Development; Drug resistance; ES Cell Line; Electroporation; Embryo; Embryo Transfer; Event; Female; Florida; Fostering; Freezing; Frequencies; Funding; Gene Expression; Gene Mutation; Gene Silencing; Gene Targeting; Gene Transfer Techniques; Generations; Genes; Genetically Engineered Mouse; Genomics; Germ Lines; Grant; HSV-Tk Gene; Haploidy; Heterozygote; Housing; Human; Human Resources; Hybrids; Injection of therapeutic agent; Interferon Type II; Journals; Karyotype determination procedure; Knock-out; Knockout Mice; Laboratories; LacZ Genes; Malignant Neoplasms; Mayo Clinic Cancer Center; Metaphase Spread; Microinjections; Molecular Cloning; Molecular Genetics; Molecular Profiling; Mothers; Mouse Strains; Mouse, Founder, Transgenic; Mus; Mutation; Nature; Neomycin; Newborn Infant; Nurses; Phase; Plasmid Cloning Vector; Plasmids; Play; Point Mutation; Positioning Attribute; Preparation; Principal Investigator; Procedures; Production; Protocols documentation; Publications; Puromycin; Regulation; Research; Research Personnel; Resource Sharing; Role; Schedule; Series; Services; Shipping; Ships; Site; Source; Southern Blotting; Staging; Superovulation; Surrogate Mothers; System; Tail; Taq Polymerase; Testing; Tetracyclines; Tissues; Transgenes; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Vertebral column; Viral; Work; base; blastocyst; cancer therapy; cell type; cohort; cost; design; egg; embryonic stem cell; experience; fusion gene; homologous recombination; hygromycin A; interest; knockout gene; male; meetings; member; mouse model; mutant; offspring; overexpression; programs; promoter; pup; research study; stem; success; tool; transgene expression; transmission process; vector; web site; zygote

Genetically engineered mice serve as a powerful tool for studying the molecular genetic basis of human cancer and for testing new cancer therapies. The main purpose of the Transgenic and Gene Targeted Mouse Shared Resource (TGTMSR) is to generate genetically altered mice for all Mayo Clinic investigators. The TGTMSR provides two main services, the generation of gene-targeted mice and the production of transgenic animals. The gene targeting service was established in 1999, while the transgenic service is a new service established in 2006 and included in this application for support. Services are very comprehensive allowing investigators with little or no prior experience in generating transgenics or knockouts to implement genetically altered mice into their research programs. We design and purify transgenic DNA constructs and inject these into fertilized eggs. We generate transgenic founders from these eggs and cross these mice to wildtype mice to transmit the transgene through the germline. Furthermore, we design gene targeting vectors (to generate classical or conditional knockouts, hypomorphic alleles or to introduce point mutations in endogenous genes), electroporate these into ES cells and screen for clones carrying the desired alteration by southern blotting. Correctly targeted clones are karyotyped and injected into host blastocysts. Chimeric males are bred to wildtype females to establish germline transmission of the targeted mutation. Our success rate of generating transgenics is thus far 100%, while that of gene targeting is >95%. All users over the current funding period have been Mayo Clinic Cancer Center (MCCC) members. Studies using mouse models generated by the TGTMSR have led to a series of scientific publications over the past 5 years, several of which were in outstanding scientific journals. A total of 27 grant applications supported by the TGTMSR were funded, 13 of which were from the NCI.

基因工程小鼠是研究人类分子遗传基础的有力工具 癌症和测试新的癌症疗法。转基因和基因靶向的主要目的 小鼠共享资源 (TGTMSR) 旨在为所有 Mayo Clinic 研究人员生成基因改造小鼠。 TGTMSR提供两项主要服务，基因靶向小鼠的产生和 转基因动物。基因打靶服务成立于1999年，而转基因服务则是 2006 年建立的新服务并包含在此支持应用程序中。服务很 全面的允许研究人员在产生转基因或基因敲除方面很少或没有经验 将基因改造小鼠纳入他们的研究计划。我们设计并纯化转基因 DNA 构建并将其注入受精卵中。我们从这些卵子中产生转基因创始人并进行杂交 将这些小鼠转化为野生型小鼠，通过种系传递转基因。此外，我们还设计了基因 靶向载体（产生经典或条件敲除、低等位基因或引入点 内源基因突变），将它们电穿孔到 ES 细胞中并筛选携带该基因的克隆 通过Southern印迹进行所需的改变。对正确靶向的克隆进行核型分析并注射到宿主中 囊胚。将嵌合雄性与野生型雌性交配以建立目标基因的种系传播 突变。迄今为止，我们转基因的成功率是 100%，而基因打靶的成功率则 >95%。 当前资助期内的所有用户均为梅奥诊所癌症中心 (MCCC) 会员。研究 使用 TGTMSR 生成的小鼠模型在过去 5 年发表了一系列科学出版物 多年，其中一些发表在杰出的科学期刊上。共有 27 项资助申请获得资助 TGTMSR 得到了资助，其中 13 项来自 NCI。