Novel method to produce transgenic rabbits

生产转基因兔的新方法

• 批准号: 6834321
• 负责人: Fuliang Du
• 金额: $ 11.48万
• 依托单位: EVERGEN BIOTECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-17 至 2006-03-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6834321
• 关键词: acute phase protein; biotechnology; cardiovascular disorder; disease /disorder model; gene targeting; genetically modified animals; laboratory rabbit; model design /development

DESCRIPTION (provided by applicant): The long term objectives of proposed study is to produce transgenic/knockout(KO) rabbits as model animals for human cardiovascular disease (CVD) and other diseases. Rabbit has been long recognized a better CVD model animal than mouse and rat, however transgenic/KO rabbit production is hampered largely by low efficiency rate of the conventional DNA microinjection method and the unavailability of embryonic stem cell lines. We propose a novel solution to aforementioned problems: use genetically manipulated somatic cells for nuclear transfer (NT) to produce transgenic/KO clones. In Phase I study, we choose C-reactive protein (CRP) as the targeted gene. We propose to produce CRP knockout cloned rabbits. The specific aims include: (1) identify 2-3 well-characterized rabbit fibroblast cell lines; (2) genetically manipulate these cell lines at different passages, generating CRP knockout cell lines; (3) produce cloned rabbit embryos using KO cell lines vs. non-KO cell lines; and (4) transfer cloned embryos to produce CRP-KO-NT rabbits. These efforts will establish the foundation for commercially producing transgenic/KO rabbits as model animals for CVD and other human disease studies. Such transgenic rabbit facility is not available in the United States at this moment and we believe our proposed project will for the first time make rabbit, the valuable however under-studied model animal, available to researchers throughout the country and worldwide.

描述（由申请方提供）：拟定研究的长期目标是生产转基因/基因敲除（KO）兔作为人类心血管疾病（CVD）和其他疾病的模型动物。兔是一种优于小鼠和大鼠的心血管疾病模型动物，但传统的DNA显微注射方法效率低，胚胎干细胞系缺乏，严重阻碍了转基因/KO兔的生产。我们提出了一种新的解决上述问题的方法：使用基因操作的体细胞进行核移植（NT），以产生转基因/KO克隆。在I期研究中，我们选择C反应蛋白（CRP）作为靶基因。我们建议生产CRP基因敲除克隆兔。具体目标包括：（1）鉴定2-3个充分表征的兔成纤维细胞系;（2）在不同传代时遗传操作这些细胞系，产生CRP敲除细胞系;（3）使用KO细胞系与非KO细胞系产生克隆兔胚胎;和（4）转移克隆胚胎以产生CRP-KO-NT兔。这些努力将为商业化生产转基因/KO兔作为CVD和其他人类疾病研究的模型动物奠定基础。这种转基因兔子设施目前在美国还没有，我们相信我们提出的项目将首次使兔子，有价值的，但研究不足的模型动物，提供给全国和世界各地的研究人员。