Is Basonuclin essential for Oogenesis

Basonuclin 对卵子发生至关重要吗

• 批准号: 6602967
• 负责人: HUNG TSENG
• 金额: $ 7.93万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6602967
• 关键词: cell proliferation; egg /ovum; gametogenesis; gene expression; gene targeting; genetic promoter element; genetic transcription; genetically modified animals; immunocytochemistry; laboratory mouse; oogenesis; polymerase chain reaction; protein biosynthesis; protein structure; recombinase; ribosomal RNA; southern blotting; spermatogenesis; transcription factor

DESCRIPTION (provided by applicant): Basonuclin, a zinc finger protein of 120 kDa, is likely to be a transcription factor that enhances transcription of ribosomal RNA (rRNA) genes. Its expression is restricted to the germ cells of the testis and ovary, and keratinocytes of the stratified epithelium. It has been long recognized that rRNA gene transcription, a key factor in determining ribosome production, is regulated according to the level of cellular protein synthesis. Certain cellular physiological states, such as proliferation (spermatogenesis) or protein accumulation (oogenesis) require a high level of protein synthesis. Although transcription factors associated with the high level of ribosome synthesis during proliferation have been described, these factors appear to be ubiquitous, whereas cell-type specific regulators of rRNA synthesis have not been reported. Basonuclin, because of its: (1) restricted cell-type distribution, (2) association with the rRNA genes in interphase as well as in mitosis, (3) interaction with the promoter of the rRNA genes, and (4) ability to increase transcription from an rDNA promtor, is likely to be a cell-type specific rRNA transcription factor, the first of its kind; (5) our attempt to knockout basonuclin gene by conventional technique was not successful because the null mutation cannot be transmitted into the germline, suggesting that basonuclin is haploinsufficient for germ cell development. Furthermore, the results of our recent study suggest that basonuclin plays a critical role during mouse oogenesis. Based on these observations, we hypothesize that basonuclin is essential in ribosome biogenesis during early mouse oogenesis. If so, inhibition, or targeted disruption, of the function of basonuclin in an oocyte should perturb its development, or the development of the embryo derived from it. Because of our earlier failure in creating a basonuelin knockout by conventional techniques, in this application, we propose, in a single Specific Aim, to test the feasibility of employing two recently developed conditional gene-disruption techniques. Sub aim 1 will use an RNAi approach to reduce the amount of basonuclin in oocytes; Sub aim 2 will use the Cre recombinase to knockout [the] basonuclin gene in oocytes.

描述（由申请人提供）：Basonuclin是120 kDa的锌指蛋白，可能是增强核糖体RNA（RRNA）基因转录的转录因子。它的表达仅限于睾丸和卵巢的生殖细胞，以及分层上皮的角质形成细胞。长期以来，人们已经认识到，根据细胞蛋白合成的水平，RRNA基因转录是确定核糖体产生的关键因素。某些细胞生理状态，例如增殖（精子发生）或蛋白质积累（卵子发生）需要高水平的蛋白质合成。尽管已经描述了与核糖体合成高水平相关的转录因子，但这些因素似乎无处不在，而尚未报道细胞类型的rRNA合成调节剂。 Basonuclin由于其：（1）限制细胞类型的分布，（2）与rRNA基因在相间和有丝分裂中的关联，（3）与rRNA基因启动子的相互作用，以及（4）增加rDNA prompor转录的能力，可能是细胞型特异性rRNA的转录因素，是第一个第一个转录因素，它的第一个转录因素，是其第一个的转录。 （5）我们试图通过常规技术敲除巴班核素基因的尝试并不成功，因为无效突变不能传输到种系中，这表明basonuclin对生殖细胞发育具有一单位倍数。此外，我们最近的研究结果表明，脑长核在小鼠卵子发生过程中起着至关重要的作用。基于这些观察结果，我们假设在小鼠早期卵子发生过程中，basonuclin在核糖体生物发生中至关重要。如果是这样，则抑制或有针对性的破坏在卵母细胞中的功能应扰动其发育，或者从中得出的胚胎的发育。由于我们较早地通过传统技术创建脊柱石蛋白敲除的失败，因此，在此应用中，我们提出了一个特定的目的，以测试采用两种最近开发的条件基因 - 局部干扰技术的可行性。 Sub Aim 1将使用RNAi方法减少卵母细胞中的盆地量； Sub AIM 2将使用CRE重组酶在卵母细胞中敲除[] Basonuclin基因。