Targeting SARS-CoV-2 RNA Pseudoknots Using Triplex-Forming Peptide Nucleic Acids

使用三链体形成肽核酸靶向 SARS-CoV-2 RNA 假结

基本信息

批准号：
10328839
负责人：
ERIKS ROZNERS
金额：
$ 19.45万
依托单位：
STATE UNIVERSITY OF NY,BINGHAMTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-11-01 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10328839
关键词：
2019-nCoV 3&apos Untranslated Regions 5&apos Untranslated Regions Affect Aminopyridines Antiviral Agents Antiviral Response Antiviral Therapy Back Binding Biological Biological Process Biology COVID-19 Calorimetry Cartoons Cations Cells Chemicals Complex Coronavirus Cultured Cells Detection Double-Stranded RNA Family Future Genetic Transcription Genetic Translation Genome Goals Health Technology Human Hydrogen Bonding Lead Life Cycle Stages Messenger RNA Molecular Conformation Mutate Mutation Nucleic Acid Binding Pathogenesis Peptide Nucleic Acids Physiological Positioning Attribute Process Protein Biosynthesis Proteins RNA RNA Conformation RNA Folding RNA Stability RNA Viruses RNA chemical synthesis Replicon Reporter Research Role SARS-CoV-2 pathogenesis Site Structure Testing Therapeutic Thermodynamics Translations Viral Viral Genome Viral Packaging Viral Physiology Viral Proteins Virus Virus Replication base combat fighting gel mobility shift assay human coronavirus innovation new therapeutic target novel novel therapeutics nucleobase pandemic disease particle pathogen peptide structure protein expression response targeted treatment tool triple helix viral RNA

项目摘要

SARS-CoV-2, the causative agent of COVID-19, is a positive-sense single-stranded RNA virus in the Coronaviridae family. The RNA of SARS-CoV-2 folds into complex double-helical structures that open the door for novel anti-viral approaches targeting RNA instead of proteins. Positive-sense RNA viruses use the specifically folded RNA structures to highjack the host cell's machinery, regulate viral replication and mRNA translation, evade antiviral responses, and produce more viral particles. The mechanisms by which SARS-CoV-2 and other coronaviruses control these processes are not well understood. However, highly conserved structural motifs of viral RNA, such as pseudoknots and hairpins, are critical for replication and translation. These motifs act as functional switches of the viral life cycle and are critical for SARS-CoV-2 pathogenesis. In this application, we propose to use triplex-forming peptide nucleic acids (PNAs) to control the conformation and biological function of SARS-CoV-2 pseudoknots. The long- term goals are to (1) better understand SARS-CoV-2 biology and pathogenesis and (2) develop new and broad-spectrum therapeutics against multiple coronavirus strains. The specific aims are to (1) study the conformational control of pseudoknot RNA switches with triplex-forming PNA and (2) study how the PNA binding and RNA switching affects coronavirus biology. We are well positioned to achieve these Aims because our research group has been working on sequence-specific recognition of biologically significant RNA molecules using triplex-forming PNAs for more than a decade. The proposed research is significant and innovative because the biology of dynamic viral RNA switches is poorly understood and have not been widely targeted in antiviral therapy. Coronaviruses can mutate their RNA genome and cross species boundaries to infect humans. Therefore, future emergence of new human coronaviruses is almost guaranteed. Mutated genomes can be rapidly sequenced, allowing us to understand how the mutations change the structure of regulatory RNA motifs. Novel therapeutic agents targeting structured RNA motifs will be applicable against multiple strains of coronaviruses and will be easily adjustable for mutated viruses, which is a key bottleneck for traditional antiviral therapeutics. If SARS-CoV-2 (or another coronavirus) mutates the PNA target site, all that is needed is to sequence the mutated genome and change the sequence of therapeutic triplex-forming PNA. RNA is emerging as a major regulatory molecule in wide variety of biological processes and as a promising novel therapeutic target. Better understanding of the biological role of viral RNA switches holds tremendous promise to result in innovative antiviral approaches to combat SARS-CoV-2 and other coronaviruses. If successful, the proposed research will develop new tools for studying the biology of complex regulatory viral RNAs, which will have broad and sustained impact on our ability to combat the current and future pandemics.