Derivation of germline competent rabbit embryonic stem cell lines

种系感受态兔胚胎干细胞系的衍生

• 批准号: 7801649
• 负责人: JIE XU
• 金额: $ 57.9万
• 依托单位: RENOVA LIFE, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-05 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7801649
• 关键词: Acute-Phase Proteins; Animal Model; Antibodies; Antibody Formation; Biochemical; C-reactive protein; Cell Line; Cells; Chimera organism; Color; Derivation procedure; Differentiation Inhibitor; Drug resistance; ES Cell Line; Embryo; Exhibits; Fibroblasts; Gene Targeting; Genes; Genetic; Germ Cells; Germ Lines; Goals; Human; Inflammation; Injection of therapeutic agent; Injury; Knock-out; Laboratories; Maintenance; Methods; Microinjections; Mus; Oryctolagus cuniculus; Pathway interactions; Phase; Plasma; Process; Production; Protocols documentation; Rattus; Recombinants; Reporting; Small Business Innovation Research Grant; Staging; Systemic infection; Tissues; Transgenic Animals; Transgenic Organisms; Work; blastocyst; drug use screening; embryonic stem cell; human ID2 protein; human disease; improved; in vivo; inhibitor/antagonist; leukemia; novel; phase 2 study; public health relevance; self-renewal; stem; success; therapeutic protein

DESCRIPTION (provided by applicant): The rabbit is a good animal model for the study of many human diseases because of its anatomical, genetic and biochemical similarities to the human. It is also used for drug screening, production of antibody and production of therapeutic proteins. The rabbit is large enough to provide adequate quantities of tissue for experimental work, produce sufficient amount of antibody or therapeutic proteins, and yet it is small enough to be economical for most laboratories. To date, pronuclear microinjection remains the primary method to produce transgenic rabbits. There are no germline competent rabbit embryonic stem (rbES) cell lines available. Pronuclear microinjection, however, is of low efficiency, results in random integration, and cannot produce gene targeted transgenic animals. The purpose of this SBIR project is to establish germline competent rbES cell lines for the production of gene targeted transgenic rabbits. In Phase I we established rabbit embryonic fibroblasts as feeder layers, synthesized recombinant rabbit leukemia inhibitory (LIF) factor, and derived putative rbES cells. In Phase II, we will work to validate if these rbES cells can contribute to germ line cells after blastocyst injection and we will work to produce gene targeted transgenic rabbits from these cells. Specific Aims in Phase II project include: (1) derive new rbES cell lines with inhibitors of differentiation; (2) evaluate germline competency of rbES cells; and (3) produce gene targeted transgenic rabbits. The success of this project will establish the first germline competent ES cell lines in a non-murine species, improve the transgenic efficiency in rabbits, and make the production of gene targeted rabbits possible. PUBLIC HEALTH RELEVANCE: The rabbit is a good animal model for the study of many human diseases because of its anatomical, genetic and biochemical similarities to the human. It is also used for drug screening, production of antibody and production of therapeutic proteins. The main reason that rabbit has not been in these fields is that there have been no established germline competent rabbit embryonic stem (rbES) cell lines. The success of this project will establish the first germline competent ES cell lines in a non-murine species, improve the transgenic efficiency in rabbits, and make the production of gene targeted rabbits possible.

描述（由申请人提供）：兔子是研究许多人类疾病的好动物模型，因为它与人的解剖学，遗传和生化相似性。它也用于药物筛查，抗体的产生和治疗蛋白的产生。兔子足够大，可以提供足够数量的组织进行实验性工作，产生足够数量的抗体或治疗蛋白，但是它足够小，可以对大多数实验室来说是经济的。 迄今为止，原核显微注射仍然是产生转基因兔子的主要方法。没有可用的生殖线既有兔胚胎茎（RBES）细胞系。然而，前核微注射效率低，导致随机整合，并且无法产生靶向基因的转基因动物。 这个SBIR项目的目的是建立生殖有能力的RBE细胞系，以生产靶向转基因兔子的基因。在第一阶段，我们建立了兔胚胎成纤维细胞为饲养层，合成的重组兔白血病抑制（LIF）因子（LIF）因子和衍生的推定RBES细胞。 在第二阶段，我们将努力验证这些RBES细胞在胚泡注射后是否可以对生殖线细胞有效，我们将努力从这些细胞中产生靶向基因的转基因兔。 II期项目中的具体目标包括：（1）带有分化抑制剂的新RBE细胞系； （2）评估RBES细胞的生殖能力； （3）产生靶向基因的转基因兔子。该项目的成功将建立在非乳腺种类中的第一个种系竞争性ES细胞系，提高兔子的转基因效率，并使靶向兔子的产生成为可能。 公共卫生相关性：兔子是许多人类疾病的良好动物模型，因为它与人的解剖学，遗传和生化相似。它也用于药物筛查，抗体的产生和治疗蛋白的产生。兔子没有进入这些领域的主要原因是没有建立的种系合格的兔胚胎茎（RBES）细胞系。该项目的成功将建立在非乳腺种类中的第一个种系竞争性ES细胞系，提高兔子的转基因效率，并使靶向兔子的产生成为可能。