A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency
We sought to explore the hypothesis that host factors required for HIV-1 replication also play a role in latency reversal. Using a CRISPR gene library of putative HIV dependency factors, we performed a screen to identify genes required for latency reactivation. We identified several HIV-1 dependency...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-08-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/9/1863 |
| _version_ | 1797576372633206784 |
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| author | Terry L. Hafer Abby Felton Yennifer Delgado Harini Srinivasan Michael Emerman |
| author_facet | Terry L. Hafer Abby Felton Yennifer Delgado Harini Srinivasan Michael Emerman |
| author_sort | Terry L. Hafer |
| collection | DOAJ |
| description | We sought to explore the hypothesis that host factors required for HIV-1 replication also play a role in latency reversal. Using a CRISPR gene library of putative HIV dependency factors, we performed a screen to identify genes required for latency reactivation. We identified several HIV-1 dependency factors that play a key role in HIV-1 latency reactivation including ELL, UBE2M, TBL1XR1, HDAC3, AMBRA1, and ALYREF. The knockout of Cyclin T1 (<i>CCNT1</i>), a component of the P-TEFb complex that is important for transcription elongation, was the top hit in the screen and had the largest effect on HIV latency reversal with a wide variety of latency reversal agents. Moreover, <i>CCNT1</i> knockout prevents latency reactivation in a primary CD4+ T cell model of HIV latency without affecting the activation of these cells. RNA sequencing data showed that CCNT1 regulates HIV-1 proviral genes to a larger extent than any other host gene and had no significant effects on RNA transcripts in primary T cells after activation. We conclude that CCNT1 function is non-essential in T cells but is absolutely required for HIV latency reversal. |
| first_indexed | 2024-03-10T21:51:14Z |
| format | Article |
| id | doaj.art-24887f62208147e084fb94f5a0c43c78 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T21:51:14Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-24887f62208147e084fb94f5a0c43c782023-11-19T13:22:47ZengMDPI AGViruses1999-49152023-08-01159186310.3390/v15091863A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from LatencyTerry L. Hafer0Abby Felton1Yennifer Delgado2Harini Srinivasan3Michael Emerman4Molecular and Cellular Biology Graduate Program, University of Washington, Seattle, WA 98195, USADivisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Center, Seattle, WA 98109, USADivisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Center, Seattle, WA 98109, USABioinformatics Shared Resource, Fred Hutchinson Cancer Center, Seattle, WA 98109, USADivisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Center, Seattle, WA 98109, USAWe sought to explore the hypothesis that host factors required for HIV-1 replication also play a role in latency reversal. Using a CRISPR gene library of putative HIV dependency factors, we performed a screen to identify genes required for latency reactivation. We identified several HIV-1 dependency factors that play a key role in HIV-1 latency reactivation including ELL, UBE2M, TBL1XR1, HDAC3, AMBRA1, and ALYREF. The knockout of Cyclin T1 (<i>CCNT1</i>), a component of the P-TEFb complex that is important for transcription elongation, was the top hit in the screen and had the largest effect on HIV latency reversal with a wide variety of latency reversal agents. Moreover, <i>CCNT1</i> knockout prevents latency reactivation in a primary CD4+ T cell model of HIV latency without affecting the activation of these cells. RNA sequencing data showed that CCNT1 regulates HIV-1 proviral genes to a larger extent than any other host gene and had no significant effects on RNA transcripts in primary T cells after activation. We conclude that CCNT1 function is non-essential in T cells but is absolutely required for HIV latency reversal.https://www.mdpi.com/1999-4915/15/9/1863HIV latencydependency factorsP-TEFb complexlatency reversal agent (LRA)CRISPR screeningCyclin T1 |
| spellingShingle | Terry L. Hafer Abby Felton Yennifer Delgado Harini Srinivasan Michael Emerman A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency Viruses HIV latency dependency factors P-TEFb complex latency reversal agent (LRA) CRISPR screening Cyclin T1 |
| title | A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency |
| title_full | A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency |
| title_fullStr | A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency |
| title_full_unstemmed | A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency |
| title_short | A CRISPR Screen of HIV Dependency Factors Reveals That <i>CCNT1</i> Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency |
| title_sort | crispr screen of hiv dependency factors reveals that i ccnt1 i is non essential in t cells but required for hiv 1 reactivation from latency |
| topic | HIV latency dependency factors P-TEFb complex latency reversal agent (LRA) CRISPR screening Cyclin T1 |
| url | https://www.mdpi.com/1999-4915/15/9/1863 |
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