Gene Targeting & Transgenic Mouse

基因打靶

基本信息

批准号：
10228635
负责人：
WINFRIED EDELMANN
金额：
$ 14.64万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-06-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10228635
关键词：
Ablation Albert Einstein Cancer Center Alleles Amino Acids Animals Bacterial Artificial Chromosomes Breeding CRISPR/Cas technology Cancer Center Cancer cell line Complex Constitutional Consult Cryopreservation DNA DNA Sequence Derivation procedure ES Cell Line Electroporation Embryo Ensure Enterobacteria phage P1 Cre recombinase Funding Gene Targeting Gene Transfer Techniques Gene-Modified Generations Genes Genomic Segment Genomics Genotype Germ Lines Human Individual Injections Knock-in Knock-in Mouse Knock-out Knockout Mice Laboratories Lentivirus Infections Location Mediating Methods Modification Mouse Strains Mus Oligonucleotides Oocytes Plasmid Cloning Vector Plasmids Price Proteins Reporter Genes Research Research Personnel Resource Sharing Risk Services Site Tamoxifen Techniques Technology Transgenes Transgenic Mice Transgenic Organisms base blastocyst conditional knockout cost efficient design design and construction embryo cryopreservation embryo stage 2 embryonic stem cell experimental study human disease human model innovation mouse genome mouse model novel novel strategies offspring preservation reproductive screening sperm cryopreservation success transgene expression vector zygote

项目摘要

PROJECT SUMMARY – GENE TARGETING AND TRANSGENIC FACILITY The Gene Targeting and Transgenic facility was founded over 28 years ago to assist AECC researchers in the generation of genetically modified mouse models of human disease. Initially, the Gene Targeting and Transgenic components operated as independent Shared Resources, however, following relocation to state-of- the-art laboratory space in the Price Center, the facilities were merged into a single Gene Targeting and Transgenic Shared Resource (GT&TF) In addition, a Gene Modification Service was implemented and integrated with the Facility to provide a complete service for the rapid and cost efficient generation of genetically modified mouse lines to all AECC investigators. The Transgenic component of the facility continues to generate, with high efficiency (~100% success rate), transgenic mouse strains through the introduction of DNA sequences, such as regular plasmid vectors or BAC clones, into the germ line by pronuclear injection or by lentivirus infection of fertilized oocytes. For each project, the facility supervisor consults with individual investigators and advises on transgene construct design, as well as making plasmids and sequence cassettes available to ensure suitability for expression in the mouse. The Gene Targeting component provided services for modification of the mouse genome through the use of CRISPR/Cas9 gene editing which was established during the current funding period. Although the vast majority of gene targeting projects are now conducted by CRISPR/Cas9, the facility continues to provide conventional gene targeting in embryonic stem (ES) cells and blastocyst injections upon request. Gene Targeting services include the generation of conventional knockout mouse lines, knock-in mouse lines and mouse lines with conditional alleles for the temporal and spatial ablation of genes. The facility has a high success rate (>95%) for obtaining gene targeted mouse lines. Finally, the services of the facility include the design and generation of simple and complex gene targeting vectors, electroporation and screening of embryonic stem cell lines as well as human and mouse cancer cell lines and the generation of chimeric mice by blastocyst injection. In addition, the facility offers mouse reproductive services for cryopreservation and re-derivation of transgenic and gene-targeted mouse lines as well as IVF services. Typically 25-30 investigators use this service each year with approximately 200 founder mice generated per year.

项目摘要——基因靶向和转基因设施基因打靶和转基因设施成立于 28 年前，旨在协助 AECC 研究人员人类疾病的转基因小鼠模型的产生最初是基因靶向和然而，在搬迁到国家后，转基因成分作为独立的共享资源运作价格中心最先进的实验室空间，这些设施被合并为一个基因靶向和转基因共享资源 (GT&TF) 此外，还实施了基因修饰服务并与该设施集成，为快速且具有成本效益的发电提供完整的服务向所有 AECC 研究人员提供转基因小鼠品系设施的转基因部分仍在继续。通过引入 DNA序列，例如常规质粒载体或BAC克隆，通过原核注射或对于每个项目，设施主管都会与个人进行协商。为研究人员提供转基因构建设计以及质粒和序列盒制作方面的建议基因靶向组件提供了服务以确保其适合在小鼠中表达。通过使用 CRISPR/Cas9 基因编辑技术修改小鼠基因组尽管目前绝大多数基因靶向项目都是由该设施继续在胚胎干 (ES) 细胞和 CRISPR/Cas9 中提供常规基因靶向根据要求提供囊胚注射服务，包括产生传统的基因敲除。小鼠品系、敲入小鼠品系以及具有时间和空间条件等位基因的小鼠品系该设施获得基因靶向小鼠品系的成功率很高（>95%）。该设施的服务包括设计和生成简单和复杂的基因靶向载体，胚胎干细胞系以及人类和小鼠癌细胞系的电穿孔和筛选通过囊胚注射产生嵌合小鼠此外，该设施还提供小鼠繁殖。转基因和基因靶向小鼠品系的冷冻保存和再衍生以及 IVF 服务通常，每年有 25-30 名研究人员使用此服务，大约有 200 只创始小鼠。每年产生。