The current COVID-19 pandemic urges the extremely sensitive and prompt detection of SARS-CoV-2 virus.Here,we present a Human Angiotensin-converting-enzyme 2(ACE2)-functionalized gold“virus traps”nanostructure as an ...The current COVID-19 pandemic urges the extremely sensitive and prompt detection of SARS-CoV-2 virus.Here,we present a Human Angiotensin-converting-enzyme 2(ACE2)-functionalized gold“virus traps”nanostructure as an extremely sensitive SERS biosensor,to selectively capture and rapidly detect S-protein expressed coronavirus,such as the current SARS-CoV-2 in the contaminated water,down to the single-virus level.Such a SERS sensor features extraordinary 106-fold virus enrichment originating from high-affinity of ACE2 with S protein as well as“virus-traps”composed of oblique gold nanoneedles,and 109-fold enhancement of Raman signals originating from multi-component SERS effects.Furthermore,the identification standard of virus signals is established by machine-learning and identification techniques,resulting in an especially low detection limit of 80 copies mL^(−1) for the simulated contaminated water by SARS-CoV-2 virus with complex circumstance as short as 5 min,which is of great significance for achieving real-time monitoring and early warning of coronavirus.Moreover,here-developed method can be used to establish the identification standard for future unknown coronavirus,and immediately enable extremely sensitive and rapid detection of novel virus.展开更多
Quantum dots（QDs）, with several unique optical and chemical features, are becoming desirable fluorescent tags for the biological applications that require long-term and highly sensitive imaging.Besides, the conjugat...Quantum dots（QDs）, with several unique optical and chemical features, are becoming desirable fluorescent tags for the biological applications that require long-term and highly sensitive imaging.Besides, the conjugation of various functional biomolecules to QDs has enabled wide applications of QDs in biological imaging. This review focuses on the following four types of QDs： semiconductor quantum dots（semiconductor QDs）, carbon nanodots（CDs）, silicon quantum dots（SiQDs）, and polymer dots（Pdots）, and summarizes the recent advancements of using these QDs in imaging microorganisms including viruses, bacteria, and fungi. We hope that this review will promote the development of new fluorescent QDs for microbial imaging and extend the applications of QD-based imaging techniques in cell biology and beyond.展开更多
Porcine deltacoronavirus(PDCoV)exploits both human aminopeptidase N(hAPN)and porcine APN(pAPN)as receptors,with a higher affinity for hAPN than for pAPN through conserved interaction sites.However,despite this affinit...Porcine deltacoronavirus(PDCoV)exploits both human aminopeptidase N(hAPN)and porcine APN(pAPN)as receptors,with a higher affinity for hAPN than for pAPN through conserved interaction sites.However,despite this affinity,PDCoV is rarely pathogenic to humans,suggesting that the utilization dynamics of APN homologs by PDCoV are distinct,which is crucial in cross-species transmission but poorly understood.Here,we employed single-virus tracking to visualize and dissect the entry dynamics of PDCoV facilitated by APN.It was discovered that APN homologs bind PDCoV simultaneously,yet the times required for the initiation of membrane fusion and internalization differ significantly.Although high-affinity hAPN,rather than low-affinity pAPN,accompanies PDCoV during internalization,low-affinity pAPN initiates PDCoV internalization approximately 20 s faster than high-affinity hAPN,with caveolae-mediated endocytosis being more predominant and productive.Depending on the cell species,low-affinity pAPN induced a 5%to 25%greater proportion and a 0.6 to 4.3 min faster cell surface fusion,contributing to efficient infection.In contrast,high-affinity hAPN resulted in a 5%to 25%greater proportion and a 5 to 15 min faster endosomal fusion,potentially promoting immune evasion.We further demonstrated that the binding affinities between the PDCoV receptor-binding domain(RBD)and APN homologs are key determinants of the differential kinetics,driving flexible transitions between the two fusion pathways.This receptor affinity-selective PDCoV entry kinetics evolves an optimal balance of immune evasion and rapid infection,underscoring the potential for PDCoV interspecies transmission and the need for its vigilant surveillance.展开更多
Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but rob...Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but robust tools to precisely elucidate the dynamic mechanisms of virus-PM fusion events are still lacking.Here,we developed a quantitative single-virus tracking approach based on highly efficient dual-color labelling of viruses and batch trajectory analysis to achieve the spatiotemporal quantification of fusion events.This approach allows us to comprehensively analyze the membrane fusion mechanism utilized by pseudotyped severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)at the singlevirus level and precisely elucidate how the relevant biomolecules synergistically regulate the fusion process.Our results revealed that SARS-CoV-2 may promote the formation of supersaturated clusters of cholesterol to facilitate the initiation of the membrane fusion process and accelerate the viral genome release.展开更多
基金the National Natural Science Foundation of China(No.51471182)this work is also supported by Shanghai international science and Technology Cooperation Fund(No.17520711700)the National Key Research and Development Project(No.2017YFB0310600).
文摘The current COVID-19 pandemic urges the extremely sensitive and prompt detection of SARS-CoV-2 virus.Here,we present a Human Angiotensin-converting-enzyme 2(ACE2)-functionalized gold“virus traps”nanostructure as an extremely sensitive SERS biosensor,to selectively capture and rapidly detect S-protein expressed coronavirus,such as the current SARS-CoV-2 in the contaminated water,down to the single-virus level.Such a SERS sensor features extraordinary 106-fold virus enrichment originating from high-affinity of ACE2 with S protein as well as“virus-traps”composed of oblique gold nanoneedles,and 109-fold enhancement of Raman signals originating from multi-component SERS effects.Furthermore,the identification standard of virus signals is established by machine-learning and identification techniques,resulting in an especially low detection limit of 80 copies mL^(−1) for the simulated contaminated water by SARS-CoV-2 virus with complex circumstance as short as 5 min,which is of great significance for achieving real-time monitoring and early warning of coronavirus.Moreover,here-developed method can be used to establish the identification standard for future unknown coronavirus,and immediately enable extremely sensitive and rapid detection of novel virus.
基金the National Natural Science Foundation of China (No. 21673037)Natural Science Foundation of Jiangsu Province (No. BK20170078)Innovative and Entrepreneurial Talent Recruitment Program of Jiangsu Province, Fundamental Research Funds for the Central Universities, and Scientific Research Foundation of Graduate School of Southeast University (No. YBJJ1778) for financial support
文摘Quantum dots（QDs）, with several unique optical and chemical features, are becoming desirable fluorescent tags for the biological applications that require long-term and highly sensitive imaging.Besides, the conjugation of various functional biomolecules to QDs has enabled wide applications of QDs in biological imaging. This review focuses on the following four types of QDs： semiconductor quantum dots（semiconductor QDs）, carbon nanodots（CDs）, silicon quantum dots（SiQDs）, and polymer dots（Pdots）, and summarizes the recent advancements of using these QDs in imaging microorganisms including viruses, bacteria, and fungi. We hope that this review will promote the development of new fluorescent QDs for microbial imaging and extend the applications of QD-based imaging techniques in cell biology and beyond.
基金supported by the National Key Research and Development Program of China(2021YFD1801104)the National Natural Science Foundation of China(32372991,32272996)+2 种基金Jiangsu Agricultural Science and Technology Innovation Fund(CX(23)1029)the Key Research and Development Program in Hunan Province,China(2023NK2017)the Fundamental Research Funds for the Central Universities(KJJQ2025021,KJYQ2025041).
文摘Porcine deltacoronavirus(PDCoV)exploits both human aminopeptidase N(hAPN)and porcine APN(pAPN)as receptors,with a higher affinity for hAPN than for pAPN through conserved interaction sites.However,despite this affinity,PDCoV is rarely pathogenic to humans,suggesting that the utilization dynamics of APN homologs by PDCoV are distinct,which is crucial in cross-species transmission but poorly understood.Here,we employed single-virus tracking to visualize and dissect the entry dynamics of PDCoV facilitated by APN.It was discovered that APN homologs bind PDCoV simultaneously,yet the times required for the initiation of membrane fusion and internalization differ significantly.Although high-affinity hAPN,rather than low-affinity pAPN,accompanies PDCoV during internalization,low-affinity pAPN initiates PDCoV internalization approximately 20 s faster than high-affinity hAPN,with caveolae-mediated endocytosis being more predominant and productive.Depending on the cell species,low-affinity pAPN induced a 5%to 25%greater proportion and a 0.6 to 4.3 min faster cell surface fusion,contributing to efficient infection.In contrast,high-affinity hAPN resulted in a 5%to 25%greater proportion and a 5 to 15 min faster endosomal fusion,potentially promoting immune evasion.We further demonstrated that the binding affinities between the PDCoV receptor-binding domain(RBD)and APN homologs are key determinants of the differential kinetics,driving flexible transitions between the two fusion pathways.This receptor affinity-selective PDCoV entry kinetics evolves an optimal balance of immune evasion and rapid infection,underscoring the potential for PDCoV interspecies transmission and the need for its vigilant surveillance.
基金supported by the National Natural Science Foundation of China(22293032,22293030,and 91859123)the National Key Research and Development Program of China(2019YFA0210500)+1 种基金the Fundamental Research Funds for the Central Universities of China(63211023)the financial support from Haihe Laboratory of Sustainable Chemical Transformations.
文摘Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but robust tools to precisely elucidate the dynamic mechanisms of virus-PM fusion events are still lacking.Here,we developed a quantitative single-virus tracking approach based on highly efficient dual-color labelling of viruses and batch trajectory analysis to achieve the spatiotemporal quantification of fusion events.This approach allows us to comprehensively analyze the membrane fusion mechanism utilized by pseudotyped severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)at the singlevirus level and precisely elucidate how the relevant biomolecules synergistically regulate the fusion process.Our results revealed that SARS-CoV-2 may promote the formation of supersaturated clusters of cholesterol to facilitate the initiation of the membrane fusion process and accelerate the viral genome release.
