EXPRESSION/FUNCTION OF C FOS AND C JUN IN DEVELOPMENT

C FOS 和 C JUN 在发育中的表达/功能

• 批准号: 3328946
• 负责人: VIRGINIA E. PAPAIOANNOU
• 金额: $ 27.94万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3328946
• 关键词: congenital disorders; developmental genetics; embryo implantation; gametogenesis; gene expression; gene mutation; genetic manipulation; genetic recombination; genetically modified animals; heterozygote; immunocytochemistry; in situ hybridization; laboratory mouse; molecular cloning; protooncogene; transposon /insertion element

The goals of this proposal are to investigate the role of proto-oncogenes in normal mammalian development and to understand how aberrations in the expression of these genes could result in pregnancy wastage or congenital malformations. C-fos and c-jun proto-oncogenes function together to regulate gene expression at the molecular level and have been chosen for this study as proto-oncogenes implicated in development. C-fos is expressed in a tissue-specific manner during embryogenesis and shows high levels of expression in extraembryonic tissues. These tissues play an essential role in implantation and in the survival of the embryo during the postimplantation period. It is our hypothesis that tissue-specific expression reflects a functional requirement for the c-fos gene product. We further postulate that c-jun will be expressed in a similar manner and will also be required during development. The specific aims of this proposal are to clarify the extent of c-fos and c-jun expression throughout the pre- and early postimplantation period in the mouse and examine the functional role of these genes in development. This will be accomplished by the following specific aims: Specific Aim I. We will detail the expression of c-fos and c-jun during pre- and postimplantation mouse development using in situ hybridization for RNA localization and immunohistochemistry for localization of the proteins. Specific Aim 2. We will produce embryonic stem (ES) cell lines heterozygous for mutated c-fos and c-jun by gene targeting via homologous recombination. Preliminary results have been successful in targeting c-fos and one mutated clone is now available. Specific Aim 3. We will then produce transgenic animals heterozygous and homozygous for mutated C-fos or c-jun by way of ES cell chimeras, in order to determine the effect of a lower dose or absence of c-fos or c-jun gene product on development. One series of chimeras is already being test-bred to produce heterozygous mice. If we determine that heterozygous cells are not capable of completing gametogenesis in a chimera, an alternative, rescue protocol will be used that will allow the study of gametogenesis in heterozygous males.

该提案的目标是研究原癌基因的作用 正常哺乳动物发育中的异常，并了解异常是如何发生的 这些基因的表达可能导致妊娠浪费或先天性畸形 畸形。 C-fos 和 c-jun 原癌基因共同发挥作用 在分子水平上调节基因表达并被选择 这项研究认为原癌基因与发育有关。 C-fos是 在胚胎发生过程中以组织特异性方式表达，并表现出高 胚胎外组织中的表达水平。这些组织起着 在胚胎着床和存活过程中发挥着重要作用 植入后时期。我们的假设是组织特异性 表达反映了c-fos基因产物的功能需求。我们 进一步假设c-jun将以类似的方式表达并且将 开发过程中也需要。 该提案的具体目的是澄清 c-fos 的范围和 c-jun 在整个植入前和植入后早期的表达 小鼠并检查这些基因在发育中的功能作用。 这将通过以下具体目标来实现： 具体目标 I。 我们将详细介绍 c-fos 和 c-jun 在预和过程中的表达。 使用 RNA 原位杂交进行植入后小鼠发育 用于蛋白质定位的定位和免疫组织化学。 具体目标2.我们将生产胚胎干（ES）细胞系 通过同源基因打靶，获得突变 c-fos 和 c-jun 的杂合子 重组。初步结果已成功靶向c-fos 现在已经可以使用一种突变克隆。 具体目标 3. 然后我们将 产生突变 C-fos 杂合子和纯合子转基因动物或 c-jun通过ES细胞嵌合体的方式，以确定a的效果 发育过程中 c-fos 或 c-jun 基因产物的剂量较低或不存在。一 系列嵌合体已经在进行试验培育以产生杂合小鼠。 如果我们确定杂合细胞无法完成 嵌合体中的配子发生，将使用替代的救援方案 这将有助于研究杂合雄性的配子发生。