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遗传物质分子遗传基础的有力工具， 癌症和测试新的癌症疗法。转基因和基因打靶的主要目的 小鼠共享资源（TGTMSR）将为所有马约诊所的研究者生产基因改变的小鼠。 TGTMSR提供两项主要服务，即基因靶向小鼠的产生和 转基因动物基因靶向服务成立于1999年，而转基因服务是一个 新服务于2006年建立，并包含在此支持申请中。服务非常 全面的允许研究者在产生转基因或基因敲除方面几乎没有或没有经验 将转基因小鼠应用到他们的研究项目中。我们设计并纯化了转基因DNA 构建并将其注入受精卵。我们从这些卵子中产生转基因创始人， 将这些小鼠转化为野生型小鼠以通过种系传递转基因。此外，我们设计基因 靶向载体（以产生经典或条件性敲除、亚纯型等位基因或引入点 内源性基因中的突变），将这些电穿孔到ES细胞中，并筛选携带内源性基因突变的克隆。 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靶向嵌合体的生殖系传播。 突变到目前为止，我们产生转基因的成功率是100%，而基因打靶的成功率是> 95%。 在当前资助期间，所有用户都是马约诊所癌症中心（MCCC）的成员。研究 使用TGTMSR生成的小鼠模型，在过去5年中发表了一系列科学出版物。 多年来，其中一些是在优秀的科学期刊。本署共批出二十七项资助申请， TGTMSR获得资助，其中13个来自NCI。