Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking

ABSTRACT Quantum dots (QDs) possess optical properties of superbright fluorescence, excellent photostability, narrow emission spectra, and optional colors. Labeled with QDs, single molecules/viruses can be rapidly and continuously imaged for a long time, providing more detailed information than when...

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Autores principales:	Li-Juan Zhang, Shaobo Wang, Li Xia, Cheng Lv, Hong-Wu Tang, Zhenpu Liang, Gengfu Xiao, Dai-Wen Pang
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	enveloped virus lipid-specific labeling quantum dot single-virus tracking Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/66e0c40c0c674f3db803f026a4f65585
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spelling	oai:doaj.org-article:66e0c40c0c674f3db803f026a4f655852021-11-15T15:56:46ZLipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking10.1128/mBio.00135-202150-7511https://doaj.org/article/66e0c40c0c674f3db803f026a4f655852020-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00135-20https://doaj.org/toc/2150-7511ABSTRACT Quantum dots (QDs) possess optical properties of superbright fluorescence, excellent photostability, narrow emission spectra, and optional colors. Labeled with QDs, single molecules/viruses can be rapidly and continuously imaged for a long time, providing more detailed information than when labeled with other fluorophores. While they are widely used to label proteins in single-molecule-tracking studies, QDs have rarely been used to study virus infection, mainly due to a lack of accepted labeling strategies. Here, we report a general method to mildly and readily label enveloped viruses with QDs. Lipid-biotin conjugates were used to recognize and mark viral lipid membranes, and streptavidin-QD conjugates were used to light them up. Such a method allowed enveloped viruses to be labeled in 2 h with specificity and efficiency up to 99% and 98%, respectively. The intact morphology and the native infectivity of viruses were preserved. With the aid of this QD labeling method, we lit wild-type and mutant Japanese encephalitis viruses up, tracked their infection in living Vero cells, and found that H144A and Q258A substitutions in the envelope protein did not affect the virus intracellular trafficking. The lipid-specific QD labeling method described in this study provides a handy and practical tool to readily “see” the viruses and follow their infection, facilitating the widespread use of single-virus tracking and the uncovering of complex infection mechanisms. IMPORTANCE Virus infection in host cells is a complex process comprising a large number of dynamic molecular events. Single-virus tracking is a versatile technique to study these events. To perform this technique, viruses must be fluorescently labeled to be visible to fluorescence microscopes. The quantum dot is a kind of fluorescent tag that has many unique optical properties. It has been widely used to label proteins in single-molecule-tracking studies but rarely used to study virus infection, mainly due to the lack of an accepted labeling method. In this study, we developed a lipid-specific method to readily, mildly, specifically, and efficiently label enveloped viruses with quantum dots by recognizing viral envelope lipids with lipid-biotin conjugates and recognizing these lipid-biotin conjugates with streptavidin-quantum dot conjugates. It is not only applicable to normal viruses, but also competent to label the key protein-mutated viruses and the inactivated highly virulent viruses, providing a powerful tool for single-virus tracking.Li-Juan ZhangShaobo WangLi XiaCheng LvHong-Wu TangZhenpu LiangGengfu XiaoDai-Wen PangAmerican Society for Microbiologyarticleenveloped viruslipid-specific labelingquantum dotsingle-virus trackingMicrobiologyQR1-502ENmBio, Vol 11, Iss 3 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	enveloped virus lipid-specific labeling quantum dot single-virus tracking Microbiology QR1-502
spellingShingle	enveloped virus lipid-specific labeling quantum dot single-virus tracking Microbiology QR1-502 Li-Juan Zhang Shaobo Wang Li Xia Cheng Lv Hong-Wu Tang Zhenpu Liang Gengfu Xiao Dai-Wen Pang Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking
description	ABSTRACT Quantum dots (QDs) possess optical properties of superbright fluorescence, excellent photostability, narrow emission spectra, and optional colors. Labeled with QDs, single molecules/viruses can be rapidly and continuously imaged for a long time, providing more detailed information than when labeled with other fluorophores. While they are widely used to label proteins in single-molecule-tracking studies, QDs have rarely been used to study virus infection, mainly due to a lack of accepted labeling strategies. Here, we report a general method to mildly and readily label enveloped viruses with QDs. Lipid-biotin conjugates were used to recognize and mark viral lipid membranes, and streptavidin-QD conjugates were used to light them up. Such a method allowed enveloped viruses to be labeled in 2 h with specificity and efficiency up to 99% and 98%, respectively. The intact morphology and the native infectivity of viruses were preserved. With the aid of this QD labeling method, we lit wild-type and mutant Japanese encephalitis viruses up, tracked their infection in living Vero cells, and found that H144A and Q258A substitutions in the envelope protein did not affect the virus intracellular trafficking. The lipid-specific QD labeling method described in this study provides a handy and practical tool to readily “see” the viruses and follow their infection, facilitating the widespread use of single-virus tracking and the uncovering of complex infection mechanisms. IMPORTANCE Virus infection in host cells is a complex process comprising a large number of dynamic molecular events. Single-virus tracking is a versatile technique to study these events. To perform this technique, viruses must be fluorescently labeled to be visible to fluorescence microscopes. The quantum dot is a kind of fluorescent tag that has many unique optical properties. It has been widely used to label proteins in single-molecule-tracking studies but rarely used to study virus infection, mainly due to the lack of an accepted labeling method. In this study, we developed a lipid-specific method to readily, mildly, specifically, and efficiently label enveloped viruses with quantum dots by recognizing viral envelope lipids with lipid-biotin conjugates and recognizing these lipid-biotin conjugates with streptavidin-quantum dot conjugates. It is not only applicable to normal viruses, but also competent to label the key protein-mutated viruses and the inactivated highly virulent viruses, providing a powerful tool for single-virus tracking.
format	article
author	Li-Juan Zhang Shaobo Wang Li Xia Cheng Lv Hong-Wu Tang Zhenpu Liang Gengfu Xiao Dai-Wen Pang
author_facet	Li-Juan Zhang Shaobo Wang Li Xia Cheng Lv Hong-Wu Tang Zhenpu Liang Gengfu Xiao Dai-Wen Pang
author_sort	Li-Juan Zhang
title	Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking
title_short	Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking
title_full	Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking
title_fullStr	Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking
title_full_unstemmed	Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking
title_sort	lipid-specific labeling of enveloped viruses with quantum dots for single-virus tracking
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/66e0c40c0c674f3db803f026a4f65585
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_version_	1718427093114552320

Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking

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