Research Training in Virus RNA Localization

病毒RNA定位研究培训

基本信息

批准号：
6794795
负责人：
CATHY L MILLER
金额：
$ 4.89万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2005-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6794795
关键词：
Reoviridae double stranded RNA genetic regulatory element genetic transcription in situ hybridization intermolecular interaction microinjections nucleic acid quantitation /detection postdoctoral investigator posttranscriptional RNA processing regulatory gene transfection virus RNA virus genetics virus protein virus replication western blottings

项目摘要

DESCRIPTION (provided by applicant): Reoviruses are prototypical members of the Reoviridae family of segmented dsRNA viruses that include important human (rotavirus) and veterinary (blue tongue disease) pathogens. The mechanisms by which 10 unique positive-sense viral RNAs are localized to viral factories for packaging into progeny core particles is poorly understood and will be addressed by the experiments in this proposal. The paths taken by reovirus positive-sense RNAs after they are transcribed will be examined by both in situ hybridization and immunostaining of bromouridine labeled viral positive-sense RNAs in infected cells. The mechanism for localization of viral RNAs to viral factories will be examined by microinjection studies of viral or control RNAs into reovirus infected cells, or transfected cells expressing reovirus RNA-binding proteins. If viral RNAs are specifically localized to viral factories, trans- and cis-acting factors involved in localization will be identified. Mechanisms for non-specific localization of RNA to factories will also be examined. Understanding the RNA localization process may lead to the development of new anti-viral strategies, and will aid in the development of a robust reverse genetics system for dsRNA viruses.

描述（由申请人提供）：呼肠孤病毒是分段 dsRNA 病毒呼肠孤病毒科的典型成员，包括重要的人类（轮状病毒）和兽医（蓝舌病）病原体。 10 种独特的正义病毒 RNA 被定位到病毒工厂并包装成后代核心颗粒的机制尚不清楚，并将通过本提案中的实验来解决。呼肠孤病毒正义RNA转录后所采取的路径将通过感染细胞中溴尿苷标记的病毒正义RNA的原位杂交和免疫染色来检查。将病毒RNA定位到病毒工厂的机制将通过将病毒或对照RNA显微注射研究到呼肠孤病毒感染的细胞或表达呼肠孤病毒RNA结合蛋白的转染细胞中来检查。如果病毒RNA特异性定位于病毒工厂，则将鉴定出参与定位的反式和顺式作用因子。还将研究 RNA 非特异性定位到工厂的机制。了解 RNA 定位过程可能会导致新的抗病毒策略的开发，并将有助于开发强大的 dsRNA 病毒反向遗传学系统。