Creation of CTIP1 and CTIP2 Null Mice

CTIP1 和 CTIP2 无效小鼠的创建

• 批准号: 6340542
• 负责人: MARK E LEID
• 金额: $ 1.88万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-29 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6340542
• 关键词: biotechnology; genetically modified animals; laboratory mouse; polymerase chain reaction; technology /technique development; tissue mosaicism

The objective of this proposal (single term, 12 months) is to create mice null for expression of COUP TF-Interacting Proteins 1 and 2 (CTIP1 and CTIP2, respectively) toward the goal of elucidating the role of each protein during embryogenesis and in the adult animal. This sabbatical period will also afford an opportunity for the PI to develop expertise in the techniques associated with genomic manipulations in the mouse including homologous recombination in and culture of embryonic stem cells, blastocyst injection and genotyping/analysis of the resultant mice. The PI's goal is to establish such technology on the campus of Oregon State University, which is currently devoid, but in great need, of a facility for creation of knock-out and transgenic animals. CTIP1 and CTIP2 represent two members of a novel family of C2H2 zinc finger proteins that were isolated in a yeast two-hybrid screen conducted to identify proteins capable of interaction with members of the COUP-TF family of orphan nuclear receptors. Both CTIP1 and CTIP2 are highly expressed in brain, as are all members of the COUP-TF family of proteins. CTIP1 has been examined in detail and was found to harbor autonomous transcriptional regulatory domains and to potentiate the transcriptional repression activity of ARP1, a COUP-TF family member, in mammalian cells in a manner that was not sensitive to reversal by the histone deacetylase inhibitor, trichostatin A. This finding suggests that transcriptional repression mediated by ARP1.CTIP1 complexes in mammalian cells may be mechanistically distinct from that of other nuclear receptors. The work described herein will facilitate elucidation of the function of the CTIP proteins in an organismal context as well as definition of the underlying molecular mechanisms by which COUP-TF family members, acting through this novel family Of C2H2 zinc finger proteins, may exert transcriptional regulatory activity during embryogenesis and in the adult animal.

该提案的目标（单期，12 个月）是创建 COUP TF 相互作用蛋白 1 和 2（分别为 CTIP1 和 CTIP2）表达无效的小鼠，以期阐明每种蛋白在胚胎发生和胚胎发育过程中的作用。成年动物。这个休假期还将为 PI 提供机会，发展与小鼠基因组操作相关的技术专业知识，包括胚胎干细胞的同源重组和培养、囊胚注射和所得小鼠的基因分型/分析。 PI 的目标是在俄勒冈州立大学校园建立此类技术，该大学目前缺乏但非常需要用于创建基因敲除和转基因动物的设施。 CTIP1 和 CTIP2 代表 C2H2 锌指蛋白新家族的两个成员，它们是在酵母双杂交筛选中分离出来的，该筛选旨在鉴定能够与孤儿核受体 COUP-TF 家族成员相互作用的蛋白质。 CTIP1 和 CTIP2 均在大脑中高表达，COUP-TF 蛋白家族的所有成员也是如此。 CTIP1 经过详细检查，发现其具有自主转录调节结构域，并以对组蛋白脱乙酰酶抑制剂的逆转不敏感的方式增强哺乳动物细胞中 COUP-TF 家族成员 ARP1 的转录抑制活性，这一发现表明，哺乳动物细胞中由 ARP1.CTIP1 复合物介导的转录抑制在机制上可能与其他核受体的转录抑制不同。本文描述的工作将有助于阐明 CTIP 蛋白在有机体环境中的功能，以及定义潜在的分子机制，COUP-TF 家族成员通过这个新的 C2H2 锌指蛋白家族发挥作用，可以发挥转录调节作用。胚胎发生期间和成年动物中的活性。