Creation of CTIP1 and CTIP2 Null Mice

本提案（单期，12个月）的目的是创建COUP TF相互作用蛋白1和2（分别为CTIP 1和CTIP 2）表达无效的小鼠，以阐明每种蛋白在胚胎发生期间和成年动物中的作用。该休假期还将为PI提供机会，以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州立大学的校园内建立这样的技术，该大学目前缺乏，但非常需要建立敲除和转基因动物的设施。CTIP 1和CTIP 2代表了C2 H2锌指蛋白新家族的两个成员，其在酵母双杂交筛选中分离，所述酵母双杂交筛选用于鉴定能够与孤儿核受体的COUP-TF家族成员相互作用的蛋白质。CTIP 1和CTIP 2在脑中高度表达，COUP-TF蛋白家族的所有成员也是如此。CTIP 1已被详细检查，发现其具有自主转录调节结构域，并以对组蛋白去乙酰化酶抑制剂阿司他丁A的逆转不敏感的方式在哺乳动物细胞中增强COUP-TF家族成员ARP 1的转录抑制活性。这一发现表明，在哺乳动物细胞中由ARP1.CTIP1复合物介导的转录抑制可能与其他核受体的机制不同。本文所述的工作将有助于阐明CTIP蛋白在生物体环境中的功能，以及定义COUP-TF家族成员通过这种新型C2 H2锌指蛋白家族发挥作用的潜在分子机制，可以在胚胎发生期间和成年动物中发挥转录调节活性。

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.