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SARS CoV-2 Nsp1 inhibitor

SARS CoV-2 Nsp1 inhibitor
SARS CoV-2 Nsp1 抑制剂
批准号：
10263809
负责人：
James Inglese
金额：
$22.11万
依托单位：
NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
--
资助国家：
美国
项目状态：
未结题
起止时间：
至
来源：
https://reporter.nih.gov/project-details/10263809
关键词：
2019-nCoVAntiviral AgentsAttenuatedBindingBiological AssayBiophysicsCell LineCellsCollaborationsColoradoGene ExpressionGenetic TranscriptionGenomicsIn VitroInterferonsLaboratoriesMutationProteinsRecombinantsReporterRibosomesSeriesSevere Acute Respiratory SyndromeSignal TransductionStructureSystemTokyoTranslationsUniversitiesValidationViral Nonstructural ProteinsVirulence FactorsVirusbasedesigninhibitor/antagonistinterestmedical schoolsresponsescreeningsite-specific integrationvaccine trialvirology

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中文摘要
翻译
qHTS检测采用了几种策略。第一，使用体外转录和翻译系统直接评估nsp1对核糖体翻译的功能影响。第二种方法包括以不同格式有条件地表达nsp1蛋白的基于细胞的试验，包括先前设计的巧合报告细胞系，该细胞系允许将任何感兴趣的蛋白（这里是SARS-CoV2 nsp1）特异性整合到包含可诱导的巧合报告系统的基因组背景中。最后，候选化合物与nsp1结合的直接生物物理评估将通过使用重组生物素化的nsp1进行基于spr的测定来验证。
英文摘要
qHTS assays have been developed using several strategies. The first, uses an in vitro transcription and translation system to directly assess the functional effect of nsp1 on ribosomal translation. The second encompass cell-based assays conditionally expressing the nsp1 protein in different formats, including a coincidence reporter cell lines previously designed to allow a site-specific integration of any protein of interest, here SARS-CoV2 Nsp1 into a genomic background containing an inducible coincidence reporter system. Finally, direct biophysical assessment of candidate compound binding to nsp1 will be verified with a SPR-based assay using recombinant biotinylated nsp1.
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