Analysis of Sense and Antisense Transcripts during the Giardia lamblia Lifecycle

贾第鞭毛虫生命周期中的正义和反义转录本分析

基本信息

批准号：
8113468
负责人：
Christopher Williams
金额：
$ 2.54万
依托单位：
GEORGETOWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-10 至 2012-05-17
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8113468
关键词：
Abdominal Pain Affect Archaea Area Azacitidine Binding Biochemical Cells Chromatin Cues Cyst DNA-Directed RNA Polymerase Dactinomycin Development Diarrhea Disease Documentation Drug usage Elements Employee Strikes Enzyme Inhibition Enzymes Eukaryota Event Flatulence Gene Expression Gene Expression Regulation Genes Genetic Transcription Genome Giardia Giardia lamblia Giardiasis Goals Grant Image In Vitro Infant Infection Intestines Lead Life Measures Mediating Membrane Proteins Messenger RNA Methods Methylation Modification Molecular Molecular Biology Nature Nonsense-Mediated Decay Parasites Parasitic Diseases Pathogenesis Pharmaceutical Preparations Phylogeny Play Plumbing Poly A Polyadenylation Positioning Attribute Post-Transcriptional Regulation Process Production Protons RNA Interference RNA Ligase (ATP)RNA Polymerase II Regulation Relative (related person)Research Research Personnel Reverse Transcriptase Polymerase Chain Reaction Ribosomal RNA Ribosomes Role Sanitation Site Specific qualifier value Staging Symptoms Transcript Translations United States Untranslated Regions Vomiting Water bisulfite chromatin remodeling drinking excystation guanylyltransferase inhibitor/antagonist interest mRNA Stability molecular chaperone GRP78 novel pathogen polyadenylated messenger RNA promoter sound transcription factor waterborne

项目摘要

DESCRIPTION (provided by applicant): This proposal is centered on understanding the transcriptional and post-transcriptional mechanisms that regulate gene control in Giardia lamblia. We are particularly interested in understanding the connection between the abundant antisense transcripts and gene expression in this intestinal protozoan pathogen. Developing a strong understanding ofthe parasite's molecular biology and gene regulation will enable researchers to develop new chemotherapeutic treatments to disrupt the critical life stages of the Giardia parasite. Our hypothesis is that the parasite uses a combination of mechanisms to regulate global gene expression during developmental periods. The research focuses on three areas of gene regulation; (1) mRNA stability, (2) chromatin-level control of gene regulation, and (3) examination of post-transcriptional modifications that affect gene expression. The specific aims are to examine the stability of sense/antisense transcripts in trophozoites and cysts, to investigate the effect of a drug that affects chromatin remodeling oh sense/antisense transcript production, and to understand the modifications and fates of mRNA sense and antisense transcripts. To measure mRNA stability, we will treat cells with the transcription inhibitor Actinomycin D and examine the rates of degradation of sense and antisense transcripts for a specified list of genes using qRT-PCR. To examine the effect of chromatin level control, specifically methylation, on transcription, we will conduct drug studies using 5-Azacytidine and quantify sense and antisense transcripts levels as above. During excystation, we will investigate the 5' modification of transcripts using RNA ligase- mediated RT-PCR. FISH will be used to image transcript localization within the cell. Giardiasis, like many other waterborne parasitic diseases is prevalent in areas that lack basic sanitation and adequate plumbing. Critical to the parasite's survival is its ability to enter and exit the cyst stage. Understanding ofthe parasite's molecular biology is a priority if we are to develop novel chemotherapeutic treatments for giardiasis.

描述（由申请人提供）：该提案的重点是了解调节贾迪亚兰布利亚基因控制的转录和转录后机制。我们特别有兴趣了解这种肠道原生动物病原体中丰富的反义转录本和基因表达之间的联系。对寄生虫的分子生物学和基因调节有深入的了解，将使研究人员能够开发新的化学治疗方法，以破坏贾第鞭毛虫的关键生命阶段。我们的假设是，寄生虫结合使用机制来调节发育时期的全球基因表达。该研究的重点是基因调节的三个领域。（1）mRNA稳定性，（2）基因调控的染色质级控制，（3）检查影响基因表达的转录后修饰。具体的目的是检查滋养体和囊肿中感觉/反义转录本的稳定性，以研究影响染色质重塑的药物的效果OH感觉/反义转录本的产生，并了解mRNA Sense和反义转录物的修改和命运。为了测量mRNA稳定性，我们将使用转录抑制剂放线霉素D治疗细胞，并检查使用QRT-PCR指定基因列表的感觉和反义转录物降解率。为了检验染色质水平对照，特异性甲基化对转录的影响，我们将使用5-氮杂丁胺进行药物研究，并量化上述感觉和反义转录物水平。在ExcyStation期间，我们将使用RNA连接酶介导的RT-PCR研究转录本的5'修饰。鱼将用于图像在细胞中的转录本定位。像许多其他水传播寄生虫疾病一样，贾第鞭毛疾病在缺乏基本卫生和足够的管道的地区普遍存在。对寄生虫的生存至关重要的是它进入和退出囊肿阶段的能力。如果我们要开发新颖的贾第鞭毛疾病化学治疗方法，那么对寄生虫的分子生物学的了解是优先的。