Transgenic & Knockout Shared Resource

转基因和基因敲除共享资源项目总结 梅奥临床癌症中心转基因与基因敲除(TKO)共享资源的主要目的是 协助所有3个地点的研究人员设计和生成转基因小鼠模型。 该资源的利用率一直很高，来自8个不同项目的49名癌症中心成员使用 本供资期间(2013-2017年)的共享资源。我们的服务包括CRISPR/CAS9 介导编辑受精卵中的感兴趣基因以产生敲除和敲入小鼠 模型使其失活或标记，或引入点突变、报告基因、loxP位点或大片段缺失。 尽管CRISPR/Cas9的基因编辑非常有用，但在某些情况下，它不像 胚胎干细胞的常规基因打靶，促使我们维持我们的传统服务 主要是插入大的(&gt；5kb)DNA片段和loxP位点。将军澳核心提供的额外服务 包括创始人筛选、创始人选育、精子冷冻保存、冷冻保存小鼠复苏 通过体外受精和标准组织病理学检测。将军澳共享资源员工亦提供 关于DNA制备、小鼠基因分型、操作和饲养的建议。保持在…的前沿 在基因组编辑方面，将军澳致力于开发或实施创新和实验 程序。上述活动允许癌症中心成员实现最先进的老鼠模型 解决癌症病因学领域的关键开放问题，并设计和测试新的治疗方法 改善癌症治疗结果的药物。将军澳共享资源在 之前的现场访问。

TRANSGENIC AND KNOCKOUT SHARED RESOURCE PROJECT SUMMARY The main purpose of the Mayo Clinic Cancer Center Transgenic and Knockout (TKO) Shared Resource is to assist investigators at all 3 sites with the design and the generation of genetically modified mouse models. Utilization of the resource has been strong with 49 Cancer Center members from 8 different Programs using the Shared Resource during the current funding period (2013-2017). Our services include CRISPR/Cas9 mediated editing of genes of interest in fertilized mouse eggs to generate knockout and knock-in mouse models to inactivate or tag it, or to introduce point mutations, reporter genes, loxP sites or large deletions. Although gene editing by CRISPR/Cas9 is incredibly useful, in some instances it is not as robust as conventional gene targeting in embryonic stem (ES) cells, prompting us to maintain our classical services mainly to insert large (>5 kb) DNA fragments and loxP sites. Additional services offered by the TKO Core include founder screening, founder breeding, sperm cryopreservation, recovery of cryopreserved mouse strains through in vitro fertilization, and standard histopathology. TKO Shared Resource staff also provide advice on DNA preparation, mouse genotyping, handling and husbandry. To stay at the cutting edge of genome editing, the TKO devotes time and effort to develop or implement innovative and experimental procedures. The above activities allow Cancer Center members to implement state-of-the-art mouse models to address key open questions in the field of cancer etiology and to design and test new therapeutic approaches that improve cancer treatment outcome. The TKO Shared Resource achieved a score of Outstanding at the previous site visit.