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（由申请人提供）：拟议研究的长期目标是生产转基因/敲除（KO）兔子作为人类心血管疾病（CVD）和其他疾病的模型动物。长期以来，兔子一直被认为是比小鼠和大鼠更好的CVD模型动物，但是转基因/KO兔的产生在很大程度上受到常规DNA微注射方法的低效率速率和胚胎干细胞系的不可用。我们提出了一种新的解决方案，以解决上述问题：使用遗传操纵的体细胞进行核转移（NT）产生转基因/KO克隆。在I期研究中，我们选择C反应蛋白（CRP）作为靶向基因。我们建议生产CRP敲除克隆的兔子。具体目的包括：（1）识别2-3个特征良好的兔成纤维细胞系； （2）遗传在不同段落处操纵这些细胞系，从而产生CRP基因敲除细胞系； （3）使用KO细胞系与非KO细胞系产生克隆的兔胚胎； （4）转移克隆的胚胎以产生CRP-KO-NT兔子。这些努力将为商业生产转基因/KO兔子作为CVD和其他人类疾病研究的模型动物的基础。这种转基因兔子设施目前尚不可用，我们相信我们提出的项目将首次制作兔子，这是有价值的模型型动物，但在全国和全球范围内可用于研究人员。