CAREER: The Mechanism and Genome-Wide Regulation of Genes and Transposable Elements by Epigenetically Active Small Interfering RNAs

智力上的功绩。转座元件(TES)是可移动的，可以插入基因，导致突变。为了防止这种突变潜力，生物进化出了抑制TES转录的机制。最近，PI的实验室发现，当TES在拟南芥植物中转录激活时，许多TE转录本被加工成大量新的小RNA，这些小RNA被称为表观遗传活性小干扰RNA(Easy RNAs)。这些Easy RNAs被认为负责启动活性TES的沉默，以DNA甲基化的形式建立表观遗传调控，以及产生大规模的转录变化。这个项目的主要目标是探索Easy RNA生物发生是如何被触发的，并了解Easy RNA生产的多种生物学功能。对这些过程的研究对于了解基因组如何保持完整性并在转录激活中幸存下来，以及细胞如何识别哪些需要表达(细胞基因)和哪些应该沉默(TES)至关重要。此外，简单RNA对多种细胞功能的影响可能解释一些重要的农业现象，因此了解这些机制以及哪些基因受到影响是基本的。该项目将支持对本科生、研究生和博士后科学家的培训和指导。此外，该项目将包括两项独立的活动，旨在最好地整合国际和平研究所的研究、教学和多样性宣传部分。首先，国际学生联合会目前的青年学者计划生物强化讲习班将继续下去，并将被用于产生青年学者计划生物学实习机会，该讲习班面向哥伦布市区公立学校系统的7年级学生。在这一新颖的计划中，STEM学科中未被充分代表的少数民族中学将被整合到皮埃尔？S实验室，在进行培训和学习练习的同时，获得真实的生物实验室经验，推进生物发现。一旦实习结束，学生将自动过渡到PI的科学指导计划。这一计划将扩大到包括其他个人投资机构，并将通过为代表不足的学生提供延长指导的专业科学研究体验来影响社区。其次，PI将把这个项目的研究完全整合到一年一度的高级本科实验室课程中。在俄亥俄州立大学教学促进中心课程设计研究所的帮助下，PI将把这个项目的具体子目标整合到本科实验室的课程和教育中。课程实验产生的原始数据将由班级在本科生研究期刊上发表，并将直接由PI使用。这门课程将利用分子遗传学系的教学基础设施来教授和培训本科生分子生物学，同时也让学生对推进科学发现感到兴奋。

Intellectual Merit. Transposable elements (TEs) are mobile and can insert into genes, causing mutations. To guard against this mutagenic potential, organisms have evolved mechanisms to repress transcription of TEs. Recently the PI's laboratory has discovered that when TEs are transcriptionally activated in Arabidopsis plants, many of the TE transcripts are processed into large quantities of novel small RNAs that are termed epigenetically active small interfering RNAs (easiRNAs). These easiRNAs are hypothesized to be responsible for the initiation of silencing of active TEs, the establishment of epigenetic regulation in the form of DNA methylation, as well as production of large-scale transcriptomic changes. The major goal of this project is to explore how easiRNA biogenesis is triggered and to understand the multiple biological functions of easiRNA production. Investigation of these processes is critical for understanding how the genome maintains integrity and survives TE transcriptional activation, as well as how a cell can recognize what needs to be expressed (cellular genes) from what should be silenced (TEs). Moreover, easiRNA influence on multiple cellular functions may explain some important agricultural phenomena, and therefore it is fundamental to understand these mechanisms and which genes are affected.Broader Impacts. This project will support training and mentoring of undergraduate, graduate and post-doctoral scientists. In addition, the project will include two independent activities designed to best integrate the PI's research, teaching, and diversity outreach components. First, the PI's current Young Scholars Program Biology Enhancement Workshop, which is taught to 7th grade students from the urban Columbus public school system, will be continued and leveraged to generate the Young Scholars Program Biology Internships. In this novel program middle school underrepresented minorities in STEM disciplines will be integrated into the PI?s laboratory and get real biology laboratory experience and advance biological discovery while engaged in training and learning exercises. Once the internship is completed, the student will automatically transition into a scientific mentoring program with the PI. This program will be expanded to include other PIs and will impact the community by providing a professional scientific research experience with extended mentoring to underrepresented students. Second, the PI will fully integrate the research in this project into an annual upper-division undergraduate laboratory course. With help from Course Design Institute of the Ohio State University Center for the Advancement of Teaching, the PI will integrate specific sub-aims from this project into the undergraduate laboratory curriculum and education. The original data produced from the course experiments will be published by the class in an undergraduate research journal and will be used directly by the PI. This course will leverage the Department of Molecular Genetics teaching infrastructure to teach and train undergraduate students molecular biology, while also exciting students about advancing scientific discovery.