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.

转基因小鼠是研究人类分子遗传学基础的有力工具 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)是为所有梅奥诊所的研究人员培育转基因小鼠。 TGTM提供两项主要服务，一是基因靶向小鼠的产生，二是转基因小鼠的生产 转基因动物。基因打靶服务成立于1999年，而转基因服务是 2006年建立的新服务，并包含在此支持申请中。服务非常好 全面的允许调查人员在产生转基因或基因敲除方面几乎或没有经验 将转基因小鼠纳入他们的研究计划。我们设计和提纯转基因DNA 构建并注射到受精卵中。我们从这些卵子中产生转基因创始人并进行杂交 这些小鼠变成野生型小鼠，通过生殖系传播转基因。此外，我们还设计了基因 靶向载体(以产生经典的或条件性的敲除、亚型等位基因或引入点 内源基因突变)，将这些基因电穿孔到ES细胞中，并筛选携带 通过Southern blotting进行所需的更改。对目标正确的克隆进行核型分析并注射到宿主体内 胚泡。将嵌合雄性繁殖到野生型雌性，以建立靶标的生殖系传播 突变。到目前为止，我们生产转基因药物的成功率是100%，而基因靶向的成功率是95%。 在当前资助期内，所有用户都是梅奥诊所癌症中心(MCCC)的成员。研究 在过去的5年里，使用由TGTM产生的小鼠模型已经导致了一系列的科学出版物 多年来，其中几篇发表在杰出的科学期刊上。共有27项拨款申请获得资助 TGTM R得到了资助，其中13个来自NCI。