Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9
The major barrier to HIV-1 cure is the persistence of latent provirus, which is not eradicated by antiretroviral therapy. The “shock and kill” approach entails stimulating viral production with latency-reversing agents followed by the killing of cells actively producing the virus by immune clearance...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-10-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/10/1154 |
| _version_ | 1827704486725943296 |
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| author | Alex Olson Binita Basukala Seunghee Lee Matthew Gagne Wilson W. Wong Andrew J. Henderson |
| author_facet | Alex Olson Binita Basukala Seunghee Lee Matthew Gagne Wilson W. Wong Andrew J. Henderson |
| author_sort | Alex Olson |
| collection | DOAJ |
| description | The major barrier to HIV-1 cure is the persistence of latent provirus, which is not eradicated by antiretroviral therapy. The “shock and kill” approach entails stimulating viral production with latency-reversing agents followed by the killing of cells actively producing the virus by immune clearance. However, this approach does not induce all intact proviruses, leaving a residual reservoir. CRISPR/Cas9 has been utilized to excise integrated Human Immunodeficiency Virus (HIV) DNA from infected cells in an RNA-guided, sequence-specific manner. Here, we seek to epigenetically silence the proviral DNA by introducing nuclease-deficient disabled Cas9 (dCas9) coupled with a transcriptional repressor domain derived from Kruppel-associated box (KRAB). We show that specific guide RNAs (gRNAs) and dCas9-KRAB repress HIV-1 transcription and reactivation of latent HIV-1 provirus. This repression is correlated with chromatin changes, including decreased H3 histone acetylation and increased histone H3 lysine 9 trimethylation, histone marks that are associated with transcriptional repression. dCas9-KRAB-mediated inhibition of HIV-1 transcription suggests that CRISPR can be engineered as a tool for block-and-lock strategies. |
| first_indexed | 2024-03-10T15:42:19Z |
| format | Article |
| id | doaj.art-5b325f9e84c34508ae8e6955b0c8362d |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T15:42:19Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-5b325f9e84c34508ae8e6955b0c8362d2023-11-20T16:45:03ZengMDPI AGViruses1999-49152020-10-011210115410.3390/v12101154Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9Alex Olson0Binita Basukala1Seunghee Lee2Matthew Gagne3Wilson W. Wong4Andrew J. Henderson5Department of Medicine and Microbiology, Boston University School of Medicine, Boston, MA 02118, USADepartment of Biology, Boston University, Boston, MA 02215, USABiomedical Engineering and Biological Design Center, Boston University, Boston, MA 02215, USADepartment of Medicine and Microbiology, Boston University School of Medicine, Boston, MA 02118, USABiomedical Engineering and Biological Design Center, Boston University, Boston, MA 02215, USADepartment of Medicine and Microbiology, Boston University School of Medicine, Boston, MA 02118, USAThe major barrier to HIV-1 cure is the persistence of latent provirus, which is not eradicated by antiretroviral therapy. The “shock and kill” approach entails stimulating viral production with latency-reversing agents followed by the killing of cells actively producing the virus by immune clearance. However, this approach does not induce all intact proviruses, leaving a residual reservoir. CRISPR/Cas9 has been utilized to excise integrated Human Immunodeficiency Virus (HIV) DNA from infected cells in an RNA-guided, sequence-specific manner. Here, we seek to epigenetically silence the proviral DNA by introducing nuclease-deficient disabled Cas9 (dCas9) coupled with a transcriptional repressor domain derived from Kruppel-associated box (KRAB). We show that specific guide RNAs (gRNAs) and dCas9-KRAB repress HIV-1 transcription and reactivation of latent HIV-1 provirus. This repression is correlated with chromatin changes, including decreased H3 histone acetylation and increased histone H3 lysine 9 trimethylation, histone marks that are associated with transcriptional repression. dCas9-KRAB-mediated inhibition of HIV-1 transcription suggests that CRISPR can be engineered as a tool for block-and-lock strategies.https://www.mdpi.com/1999-4915/12/10/1154HIV transcriptionHIV latencyCRISPRchromatin |
| spellingShingle | Alex Olson Binita Basukala Seunghee Lee Matthew Gagne Wilson W. Wong Andrew J. Henderson Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9 Viruses HIV transcription HIV latency CRISPR chromatin |
| title | Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9 |
| title_full | Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9 |
| title_fullStr | Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9 |
| title_full_unstemmed | Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9 |
| title_short | Targeted Chromatinization and Repression of HIV-1 Provirus Transcription with Repurposed CRISPR/Cas9 |
| title_sort | targeted chromatinization and repression of hiv 1 provirus transcription with repurposed crispr cas9 |
| topic | HIV transcription HIV latency CRISPR chromatin |
| url | https://www.mdpi.com/1999-4915/12/10/1154 |
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