Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release
Abstract Background The continued persistence of HIV-1 as a public health concern due to the lack of a cure calls for the development of new tools for studying replication of the virus. Here, we used NanoLuc, a small and extremely bright luciferase protein, to develop an HIV-1 bioluminescent reporte...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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BMC
2020-05-01
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| Series: | Retrovirology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12977-020-00521-5 |
| _version_ | 1818980936564342784 |
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| author | James Kirui Eric O. Freed |
| author_facet | James Kirui Eric O. Freed |
| author_sort | James Kirui |
| collection | DOAJ |
| description | Abstract Background The continued persistence of HIV-1 as a public health concern due to the lack of a cure calls for the development of new tools for studying replication of the virus. Here, we used NanoLuc, a small and extremely bright luciferase protein, to develop an HIV-1 bioluminescent reporter virus that simplifies functional measurement of virus particle production. Results The reporter virus encodes a Gag protein containing NanoLuc inserted between the matrix (MA) and capsid (CA) domains of Gag, thereby generating virus particles that package high levels of the NanoLuc reporter. We observe that inserting the NanoLuc protein within HIV-1 Gag has minimal impact on Gag expression and virus particle release. We show that the reporter virus recapitulates inhibition of HIV-1 particle release by Gag mutations, the restriction factor tetherin, and the small-molecule inhibitor amphotericin-B methyl ester. Conclusion These results demonstrate that this vector will provide a simple and rapid tool for functional studies of virus particle assembly and release and high-throughput screening for cellular factors and small molecules that promote or inhibit HIV-1 particle production. |
| first_indexed | 2024-12-20T17:23:21Z |
| format | Article |
| id | doaj.art-57d968b8f48e4e47a37ccd73b192f262 |
| institution | Directory Open Access Journal |
| issn | 1742-4690 |
| language | English |
| last_indexed | 2024-12-20T17:23:21Z |
| publishDate | 2020-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Retrovirology |
| spelling | doaj.art-57d968b8f48e4e47a37ccd73b192f2622022-12-21T19:31:42ZengBMCRetrovirology1742-46902020-05-0117111110.1186/s12977-020-00521-5Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus releaseJames Kirui0Eric O. Freed1Virus-Cell Interaction Section, HIV Dynamics and Replication Program, Center for Cancer Research, National Cancer InstituteVirus-Cell Interaction Section, HIV Dynamics and Replication Program, Center for Cancer Research, National Cancer InstituteAbstract Background The continued persistence of HIV-1 as a public health concern due to the lack of a cure calls for the development of new tools for studying replication of the virus. Here, we used NanoLuc, a small and extremely bright luciferase protein, to develop an HIV-1 bioluminescent reporter virus that simplifies functional measurement of virus particle production. Results The reporter virus encodes a Gag protein containing NanoLuc inserted between the matrix (MA) and capsid (CA) domains of Gag, thereby generating virus particles that package high levels of the NanoLuc reporter. We observe that inserting the NanoLuc protein within HIV-1 Gag has minimal impact on Gag expression and virus particle release. We show that the reporter virus recapitulates inhibition of HIV-1 particle release by Gag mutations, the restriction factor tetherin, and the small-molecule inhibitor amphotericin-B methyl ester. Conclusion These results demonstrate that this vector will provide a simple and rapid tool for functional studies of virus particle assembly and release and high-throughput screening for cellular factors and small molecules that promote or inhibit HIV-1 particle production.http://link.springer.com/article/10.1186/s12977-020-00521-5Reporter virusNano-luciferaseVirus release |
| spellingShingle | James Kirui Eric O. Freed Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release Retrovirology Reporter virus Nano-luciferase Virus release |
| title | Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release |
| title_full | Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release |
| title_fullStr | Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release |
| title_full_unstemmed | Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release |
| title_short | Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release |
| title_sort | generation and validation of a highly sensitive bioluminescent hiv 1 reporter vector that simplifies measurement of virus release |
| topic | Reporter virus Nano-luciferase Virus release |
| url | http://link.springer.com/article/10.1186/s12977-020-00521-5 |
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