Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion
Summary: Site-specific fluorescent labeling of HIV-1 proteins enables direct visualization of their dynamics and trafficking. Here, we present a protocol to label HIV-1 envelope (Env) glycoproteins on virions using genetic code expansion and click chemistry. We describe steps for using HEK293T cells...
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| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725004010 |
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| author | Wang Xu Narendra Kumar Gonepudi Maolin Lu |
| author_facet | Wang Xu Narendra Kumar Gonepudi Maolin Lu |
| author_sort | Wang Xu |
| collection | DOAJ |
| description | Summary: Site-specific fluorescent labeling of HIV-1 proteins enables direct visualization of their dynamics and trafficking. Here, we present a protocol to label HIV-1 envelope (Env) glycoproteins on virions using genetic code expansion and click chemistry. We describe steps for using HEK293T cells to produce amber-free HIV-1 carrying a noncanonical amino acid (ncAA) and for fluorophore conjugation to the ncAA. This protocol offers a practical way to apply amber suppression and click chemistry to track viral proteins during the infection lifecycle.For complete details on the use and execution of this protocol, please refer to Ao et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-ab5c03f70fbd4fbf9cabd4664f60f388 |
| institution | DOAJ |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-ab5c03f70fbd4fbf9cabd4664f60f3882025-08-20T02:48:05ZengElsevierSTAR Protocols2666-16672025-09-016310399510.1016/j.xpro.2025.103995Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansionWang Xu0Narendra Kumar Gonepudi1Maolin Lu2Department of Cellular and Molecular Biology, School of Medicine, University of Texas at Tyler Health Science Center, Tyler, TX 75708, USA; Corresponding authorDepartment of Cellular and Molecular Biology, School of Medicine, University of Texas at Tyler Health Science Center, Tyler, TX 75708, USA; Corresponding authorDepartment of Cellular and Molecular Biology, School of Medicine, University of Texas at Tyler Health Science Center, Tyler, TX 75708, USA; Corresponding authorSummary: Site-specific fluorescent labeling of HIV-1 proteins enables direct visualization of their dynamics and trafficking. Here, we present a protocol to label HIV-1 envelope (Env) glycoproteins on virions using genetic code expansion and click chemistry. We describe steps for using HEK293T cells to produce amber-free HIV-1 carrying a noncanonical amino acid (ncAA) and for fluorophore conjugation to the ncAA. This protocol offers a practical way to apply amber suppression and click chemistry to track viral proteins during the infection lifecycle.For complete details on the use and execution of this protocol, please refer to Ao et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725004010Cell-based AssaysMicrobiologyChemistry |
| spellingShingle | Wang Xu Narendra Kumar Gonepudi Maolin Lu Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion STAR Protocols Cell-based Assays Microbiology Chemistry |
| title | Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion |
| title_full | Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion |
| title_fullStr | Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion |
| title_full_unstemmed | Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion |
| title_short | Protocol for click labeling of HIV-1 envelope on amber-free virions prepared using genetic code expansion |
| title_sort | protocol for click labeling of hiv 1 envelope on amber free virions prepared using genetic code expansion |
| topic | Cell-based Assays Microbiology Chemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725004010 |
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