Epigenetic Regulation of Gene Expression During Early Mouse Embryogenesis

小鼠早期胚胎发生过程中基因表达的表观遗传调控

• 批准号: 7333934
• 负责人: M MITCHELL SMITH
• 金额: $ 5.89万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-22 至 2008-08-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7333934
• 关键词: Acetylation; Address; Alleles; Animal Model; Assisted Reproductive Techniques; Automobile Driving; Bioinformatics; Biological Assay; Biology; Cells; Centromere; Chromatin; Class; Collaborations; Commit; Complex; DNA biosynthesis; Development; Developmental Gene; Embryo; Embryonic Development; Engineering; Ensure; Enzymes; Epigenetic Process; Experimental Designs; Experimental Models; Family; Fellowship; Foundations; Future; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Genome; Goals; Health; Histone Acetylation; Histone H4; Histones; Human; Individual; Knock-out; Laboratories; Messenger RNA; Microscopy; Mind; Modification; Molecular Genetics; Mouse Strains; Mus; Nuclear; Nucleosomes; Oocytes; Pathway interactions; Pattern; Play; RNA Interference; Range; Reporter Genes; Research; Research Personnel; Research Training; Residencies; Resolution; Role; Saccharomyces cerevisiae; Saccharomycetales; Staging; Stream; Structure; Techniques; Testing; Thinking; Time; Training; Training Programs; Transgenic Organisms; Variant; Yeasts; chromatin immunoprecipitation; design; embryonic stem cell; histone acetyltransferase; interest; nuclear transfer; programs; promoter; recombinase; repaired; research study; stem cell therapy; success

DESCRIPTION (provided by applicant): One of the most remarkable gene reprogramming senarios in all of biology is the transformation of the transcriptionally silent fully grown oocyte into the totipotent embryonic stem cell. Epigenetic regulatory factors, including histone modifications and nucleosome remodeling complexes, play essential roles in this oocyte-to-embryo transition (OET). However, little is known about the specific factors involved, the program of histone modifications required for normal repropramming, or the modifications that occur at individual down-stream target genes. For over 25 years, my laboratory has used the budding yeast, Saccharomyces cerevisiae, as a model organism for dissecting the roles of histone modifications and variants in mRNA transcription, DNA replication and repair, and centromere function. Recently, our studies have expanded to focus on histone H4 acetylation by Myst2 in the preimplantation mouse embryo. Results from these preliminary experiments suggest that Myst2 is the enzyme responsible for H4 acetylation in reprogramming and zygotic gene activation. This is a major change in my research direction and success will require advanced expertise in mouse molecular genetics. To accomplish this goal, I have arranged to take a sabbatical in the laboratory of Dr. Barbara Knowles at the Jackson Laboratory. This Kirschstein-NRSA Senior Fellowship will enable a full year's residency, rather than six months, and ensure a complete training program in techniques and approaches that cannot be completed in a short tenure. During my sabbatical, I propose to address three specific aims. First, we will determine the pattern of histone modifications during OET, globally by high resolution 4Pi microscopy, and specifically at key reporter genes identified by the transcriptome analysis of the Knowles group. Second, we will engineer transgenic and knockout mouse strains that permit maternal depletion of Myst2 expression in the oocyte. We will use these strains to test directly for the requirement of Myst2 during OET. Third, we will examine the fate of H4 acetylation in Myst2- depleted embryos during OET, specifically at reporter genes and globally using tiled promoter microarrays. The results of these studies will have a significant impact on how we think about, and manage, a wide range of human health issues including assisted reproductive techniques, somatic nuclear transfer, and stem cell therapy.

描述（由申请人提供）：所有生物学中最引人注目的基因重编程之一是转录静音的完全生长的卵母细胞转化为卵黄胚胎干细胞。表观遗传调节因子，包括组蛋白的修饰和核小体重塑络合物，在这种卵母细胞到孔晶过渡（OET）中起着至关重要的作用。但是，对所涉及的特定因素，正常重启动所需的组蛋白修饰程序或在单个下游靶基因上发生的修改知之甚少。 25年来，我的实验室一直使用酿酒酵母酿酒酵母作为模型生物，用于剖析组蛋白修饰和变体在mRNA转录，DNA复制和修复中的作用，以及中心功能。最近，我们的研究已扩展到植入前小鼠胚胎中的MYST2专注于组蛋白H4乙酰化。这些初步实验的结果表明，MYST2是负责重编程和合子基因激活中H4乙酰化的酶。这是我的研究方向的重大变化，成功将需要小鼠分子遗传学方面的高级专业知识。为了实现这一目标，我已经安排在杰克逊实验室的芭芭拉·诺尔斯博士的实验室中休假。这项Kirschstein-NRSA高级奖学金将使整整一年的居留权而不是六个月，并确保在短短任期内无法完成的技术和方法进行完整的培训计划。在休假期间，我建议解决三个特定目标。首先，我们将通过高分辨率4PI显微镜，特别是通过Knowles组的转录组分析确定的关键报告基因，在全球范围内确定组蛋白修饰的模式。其次，我们将设计允许卵母细胞中MYST2表达的母体耗竭的转基因和敲除小鼠菌株。我们将使用这些菌株直接测试OET期间MYST2的需求。第三，我们将在OET期间检查MyST2耗尽的胚胎中H4乙酰化的命运，特别是使用瓷砖启动子微阵列在报告基因和全球范围内。这些研究的结果将对我们的思考和管理方式产生重大影响，包括辅助生殖技术，体细胞核转移和干细胞疗法。