Small RNA Discovery and Analysis in Ascaris

蛔虫中小 RNA 的发现和分析

基本信息

批准号：
7530994
负责人：
RICHARD E. DAVIS
金额：
$ 24.87万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-06-15 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7530994
关键词：
Address Area Ascaris Biochemical Bioinformatics Biological Biological Models Biology Cell physiology Cell-Free System Cells Child Chromatin Chromosomes Class Cognitive Condition DNA DNA Sequence Rearrangement Data Data Set Depth Developing Countries Development Drug Delivery Systems Economics Embryo Event Gametogenesis Gene Expression Gene Expression Profile Gene Expression Regulation Gene Silencing Genetic Transcription Genetic Translation Genome Goals Healed Helminths Heterochromatin Individual Intestines Knowledge Larva Lead Libraries Life Location Maintenance Methods Modeling Modification Molecular Morbidity - disease rate Muscle Nematoda Numbers Organ Organogenesis Parasites Parasitic infection Parasitic nematode Pattern Pharyngeal structure Pilot Projects Play Process Public Health RNA RNA Interference RNA Splicing RNA analysis RNA library Reading Regulation Research Role Role playing therapy Running Small RNA Social Development Staging System Systems Analysis Technology Testing Tetrahymena Tissues Transfection Translations female reproductive system healing improved in vivo male novel novel therapeutics parasitism reproductive size socioeconomics tool

项目摘要

DESCRIPTION (provided by applicant): Parasitic nematodes remain a significant public health problem in many parts of the world. Ascaris infects over 1 billion people, decreases physical and cognitive development in children, and hinders socioeconomic development in endemic areas. In order to help combat these parasitic infections, greater knowledge of parasite biology is needed. One new avenue of research yet to be explored in parasitic nematodes is small RNAs that act as "guides" in gene silencing. Small RNAs (19-32 nt) regulate mRNA translation and stability, lead to gene silencing through the formation of heterochromatin, and are involved in DNA elimination and rearrangements. Nothing is known regarding the spectrum of small RNAs present in parasitic nematodes. Ascaris represents a unique nematode system for analysis of small RNAs because 1) large numbers of synchronously developing embryos can be obtained, 2) individual tissues with discrete developmental regions such as the male and female reproductive systems can be isolated, and 3) an unusual form of DNA elimination (chromatin diminution in which 25% of the genome is eliminated) in the somatic lineages occurs during early development. We will prepare and "deep sequence" Ascaris small RNA libraries from several early developmental stages and regions of the male and female reproductive systems. Sequences from the small RNA libraries will enable us to test the following hypotheses: 1) Expression of a discrete set of small RNAs is temporally associated with chromatin diminution in early Ascaris embryos, 2) Small RNAs are differentially expressed and are temporally associated with regulation of mRNA translation and stability prior to the onset of zygotic transcription and in early development, and 3) Small RNAs are uniquely associated with the germline, gametogenesis, and organogenesis. Furthermore, we will also initiate studies to develop methods for in depth mechanistic analyses of the role of small RNAs in mRNA translation and stability in Ascaris using our cell-free and in vivo systems. Our studies will identify and characterize parasitic nematode small RNAs in specific stages and tissues and at a depth heretofore not previously done in a nematode. Overall, our analysis of small RNAs in Ascaris, a parasitic nematode with tremendous reproductive capacity, may identify novel small RNAs associated with parasitic nematodes. These studies are likely to identify new players in parasitic nematode gene expression and regulation that could be new drug targets for the development of novel therapeutics against these parasites. PUBLIC HEALTH RELEVANCE: Parasitic nematodes remain a significant public health problem in many parts of the world. Ascaris alone infects upwards of 1 billion people and hinders socioeconomic development in endemic areas. These studies are likely to identify new players in parasitic nematode gene expression and regulation that could lead to the development of novel therapeutics against parasitic nematodes.

描述（由申请人提供）：寄生线虫在世界许多地区仍然是一个重大的公共卫生问题。蛔虫感染超过 10 亿人，降低儿童的身体和认知发育，并阻碍流行地区的社会经济发展。为了帮助对抗这些寄生虫感染，需要更多的寄生虫生物学知识。寄生线虫中尚未探索的一项新研究途径是在基因沉默中充当“向导”的小RNA。小 RNA (19-32 nt) 调节 mRNA 翻译和稳定性，通过异染色质的形成导致基因沉默，并参与 DNA 消除和重排。关于寄生线虫中存在的小 RNA 谱，我们一无所知。蛔虫代表了一种用于分析小 RNA 的独特线虫系统，因为 1) 可以获得大量同步发育的胚胎，2) 可以分离具有离散发育区域的个体组织，如雄性和雌性生殖系统，3) 一种不寻常的形式体细胞谱系中的 DNA 消除（染色质减少，其中 25% 的基因组被消除）发生在早期发育过程中。我们将从男性和女性生殖系统的几个早期发育阶段和区域制备蛔虫小RNA文库并进行“深度测序”。小 RNA 文库的序列将使我们能够检验以下假设：1) 一组离散小 RNA 的表达与早期蛔虫胚胎中染色质的减少暂时相关，2) 小 RNA 存在差异表达，并且与染色质的调节暂时相关。合子转录开始前和早期发育中的 mRNA 翻译和稳定性，3) 小 RNA 与种系、配子发生和器官发生独特相关。此外，我们还将开展研究，开发使用我们的无细胞和体内系统对蛔虫中小 RNA 在 mRNA 翻译和稳定性中的作用进行深入机制分析的方法。我们的研究将鉴定和表征特定阶段和组织中的寄生线虫小RNA，其深度此前从未在线虫中进行过。总体而言，我们对蛔虫（一种具有巨大繁殖能力的寄生线虫）中的小 RNA 进行分析，可能会鉴定出与寄生线虫相关的新型小 RNA。这些研究可能会发现寄生线虫基因表达和调控的新参与者，这些参与者可能成为开发针对这些寄生虫的新疗法的新药物靶点。公共卫生相关性：寄生线虫在世界许多地区仍然是一个重大的公共卫生问题。仅蛔虫就感染了超过 10 亿人，并阻碍了流行地区的社会经济发展。这些研究可能会发现寄生线虫基因表达和调控的新参与者，从而开发针对寄生线虫的新疗法。