Transgenic

转基因

基本信息

批准号：
8382241
负责人：
Jan M. van Deursen
金额：
$ 16.21万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8382241
关键词：
Alleles Animal Model Animals Applications Grants Arizona Autologous Biological Assay Biopsy Breeding Cell division Cells Characteristics Chimera organism Clinic Collection Complementary DNA Comprehensive Cancer Center Consultations DNA DNA Sequence Rearrangement Development Embryo Embryo Transfer Female Florida Fostering Funding Gene Expression Gene Mutation Gene Targeting Gene Transfer Techniques Generations Genes Genetically Engineered Mouse Genomics Germ Lines Grant Haploidy Housing Human Human Resources Hybrids Injection of therapeutic agent Interferon Type II Journals Knock-out Knockout Mice Malignant Neoplasms Mayo Clinic Cancer Center Microinjections Molecular Cloning Molecular Genetics Molecular Profiling Mothers Mouse Strains Mouse, Founder, Transgenic Mus Mutation Nature Newborn Infant Nurses Plasmid Cloning Vector Plasmids Play Point Mutation Positioning Attribute Preparation Procedures Production Protocols documentation Publications Regulation Research Research Personnel Resource Sharing Role Series Services Shipping Ships Site Southern Blotting Staging Superovulation Surrogate Mothers System Tail Testing Tetracyclines Tissues Transgenes Transgenic Animals Transgenic Mice Transgenic Organisms Vertebral column Viral Work base blastocyst cancer therapy cohort design egg embryonic stem cell experience homologous recombination interest male meetings member mouse model mutant offspring overexpression programs promoter pup success tool transgene expression transmission process vector web site zygote

项目摘要

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强大工具癌症和测试新的癌症疗法。靶向转基因和基因的主要目的小鼠共享资源（TGTMSR）是为所有梅奥诊所研究者生成遗传改变的小鼠。 TGTMSR提供了两种主要服务，生成基因靶向的小鼠和生产转基因动物。基因靶向服务于1999年成立，而转基因服务是新服务于2006年成立，并包括在本申请中的支持中。服务非常全面允许几乎没有或没有生成转基因或淘汰的研究人员将遗传变化的小鼠实施到他们的研究计划中。我们设计和纯化转基因DNA 构造并将其注入受精卵。我们从这些鸡蛋中产生转基因创始人，然后交叉这些小鼠到野生型小鼠以通过种系传输转基因。此外，我们设计基因靶向向量（生成经典或有条件的敲除，造型等位基因或引入点内源基因的突变），将它们电摄取到ES细胞中，并筛选出携带的克隆南方印迹所需的改变。正确靶向的克隆被核分型并注入主机胚泡。嵌合雄性繁殖到野生型雌性，以建立目标的种系传播突变。到目前为止，我们产生转基因的成功率是100％，而基因靶向的成功率> 95％。当前资金期间的所有用户都是Mayo临床癌症中心（MCCC）成员。研究使用TGTMSR生成的鼠标模型在过去的5个中导致了一系列科学出版物几年，其中一些是在杰出的科学期刊中。共有27个赠款申请由 TGTMSR资助，其中13个来自NCI。