"Characterization of the germ plasm of the WASP Nasonic Vitripennis"

描述（由申请人提供）：本提案的主要目的是提供一个完整的表征的RNA组成的黄蜂的种质，然后开始功能测试确定的因素定位于该细胞器的过程。了解构成这种结构的分子是很重要的，因为种质具有独特的能力，可以推动幼稚核迅速进入种系发育程序。原始生殖细胞的许多特征对发育和疾病都有普遍的意义，包括一般转录静止期、多能性潜能和跨上皮迁移行为。纳索尼亚的种质是特别感兴趣的，因为它是一个异常大的、离散的核糖核蛋白实体，与其他种质的小颗粒特征相反。此外，纳索尼亚的原始生殖细胞的行为与果蝇的不同，表明该实体在黄蜂中的分子组成不同。已建立的从果蝇中分离胚胎片段的技术将应用于纳索尼亚胚胎。利用定量RNA测序技术，将种质物质高度富集的片段与相应的物质贫化片段直接进行比较。那些转录本在种质中含有的部分与被耗尽的部分相比被显著地过度代表，将成为种质的候选成分。这些将通过原位杂交和RNAi功能测试来证实。本研究将涉及发育生物学、RNA调控、细胞生物学、基因组学和进化生物学等领域。这项工作将包括一名研究生的强烈参与，他将在明年被招募，也可能涉及一名本科生。预计这些学生将处于早期培训水平的经验，首席研究员将投入大量时间来培训和指导招募的学生。

DESCRIPTION (provided by applicant): The main objective of this proposal is to provide a complete characterization of the RNA composition germ plasm of the wasp Nasonia vitripennis, and to then to begin the process of functionally testing confirmed factors localized to this organelle. Understanding the molecules that make up this structure is important since germ plasm has the unique capability to drive naive nuclei rapidly into the germline developmental program. Many of the characteristics of primordial germ cells are of general interest to both development and disease, including periods of general transcriptional quiescence, the potential for pluripotency, and trans-epithelial migratory behavior. The Nasonia germ plasm is of particular interest, as it is an unusually large, discrete ribonucleoprotein entity, in contrast tothe smaller granules characteristic of other germ plasms. In addition, the primordial germ cells of Nasonia behave differently from those of Drosophila, indicating a divergent molecular make-up of this entity in the wasp. Established techniques for isolating embryo fragments from Drosophila will be applied to Nasonia embryos. Fragments highly enriched for germ plasm material will be compared directly to corresponding fragments depleted for the material, by using quantitative RNA sequencing technologies. Those transcripts that are significantly overrepresented in the germ plasm containing fraction compared to the depleted fraction will be candidate components of the germ plasm. These will be confirmed by in situ hybridization, and by functional testing with RNAi. This study will touch on the fields of developmental biology, RNA regulation, cell biology, genomics, and evolutionary biology. This work will include the intense participation of a graduate student, who will be recruited in the coming year, and may also involve an undergraduate student. It is expected that the students will be at the early training level of experience, and the Principal Investigator will invest significant time in trainig and mentoring the recruited student(s).