Zinc-finger-nucleases mediate specific and efficient excision of HIV-1 proviral DNA from infected and latently infected human T cells
Open accessAbstract
HIV-infected individuals currently cannot be completely cured because existing antiviral therapy regimens do not address HIV provirus DNA, flanked by long terminal repeats (LTRs), already integrated into host genome. Here, we present a possible alternative therapeutic approach to specifically and directly mediate deletion of the integrated full-length HIV provirus from infected and latently infected human T cell genomes by using specially designed zinc-finger nucleases (ZFNs) to target a sequence within the LTR that is well conserved across all clades. We designed and screened one pair of ZFN to target the highly conserved HIV-1 5'-LTR and 3'-LTR DNA sequences, named ZFN-LTR. We found that ZFN-LTR can specifically target and cleave the full-length HIV-1 proviral DNA in several infected and latently infected cell types and also HIV-1 infected human primary cells in vitro. We observed that the frequency of excision was 45.9% in infected human cell lines after treatment with ZFN-LTR, without significant host-cell genotoxicity. Taken together, our data demonstrate that a single ZFN-LTR pair can specifically and effectively cleave integrated full-length HIV-1 proviral DNA and mediate antiretroviral activity in infected and latently infected cells, suggesting that this strategy could offer a novel approach to eradicate the HIV-1 virus from the infected host in the future.
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Various gene editing technologies have been shown to remove proviral HIV-1 DNA from the host cell genome by targeting its highly conserved 5′ and 3′ long terminal repeats.
“various gene editing technologies have recently been shown to remove the proviral HIV-1 DNA from the host cell genome by targeting its highly conserved 5′ and 3′ long terminal repeats (LTRs)”
From RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection (2014) · cites this paper as [13]
The proof of concept is applicable to any other cell type, including T-lymphoid cells, astrocytes, and neural stem cells.
“However, this proof of concept is certainly applicable to any other cell type, including T-lymphoid cells (Fig. S6) (12,13), astrocytes, and neural stem cells”
From RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection (2014) · cites this paper as [13]
It has been recognized that targeting latent infections with viral genome-specific nucleases would be useful, but engineering sequence-specific nucleases has hampered progress.
“It has been recognized that it would be useful to target latent infections with viral genome-specific nucleases (3-5), but the challenges of engineering sequence-specific nucleases have hampered progress.”
From RNA-guided endonuclease provides a therapeutic strategy to cure latent herpesviridae infection (2014) · cites this paper as [4]
Disruption of either CCR5 or CXCR4 opens up the possibility of generating alternative co-receptor using mutants.
“Moreover, disruption of either gene opens up the possibility of generating alternative co-receptor using mutants [33].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [33]
Most publications on HIV-1 gene therapy by ZFNs have focused on co-receptor CCR5 or CXCR4 disruption, but targeting the HIV-1 genome is a possible alternative strategy.
“As for HIV-1 gene therapy by ZFNs, most publications have focused on co-receptor CCR5 or CXCR4 disruption [30,31,32], but targeting HIV-1 genome is a possible alternative strategy [33].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [33]
Proof-of-principle preliminary studies demonstrate that targeting the provirus is a potential strategy for curing HIV infection.
“Nevertheless, the proof-of-principle preliminary studies described above demonstrate that targeting the provirus is a potential strategy for curing HIV infection.”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [33]
The Zhu group used a specially designed ZFN to target the highly conserved 5′ and 3′ LTR sequences of HIV-1 to successfully eradicate HIV-1 proviral DNA in cell lines as well as in acutely infected and latently infected primary T cells.
“Most recently, the Zhu group in China used a specially designed ZFN to target the highly conserved 5′ and 3′ LTR sequences of HIV-1 to successfully eradicate HIV-1 proviral DNA in cell lines as well as in acutely infected and latently infected primary T cells [33].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [33]
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- Authors
- Qu, X., Wang, P., Ding, D., Li, L., Wang, H., Ma, L., Zhou, X., Liu, S., Lin, S., Wang, X., Zhang, G., Liu, S., Liu, L., Wang, J., Zhang, F., Lu, D., & Zhu, H.
- Affiliations
- Chinese Academy of Sciences (CN)
- Corresponding author
- Zhu, Huanzhang
- ORCID
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2013-06-26
- Volume
- 41
- Issue
- 16
- Pages
- 7771-7782
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC3763554
- OpenAlex
- W1964761653
- Cited by
- 155
- References
- 41
- Citations by year
- 2026: 4 · 2025: 3 · 2024: 8 · 2023: 5 · 2022: 5 · 2021: 8 · 2020: 14 · 2019: 8 · 2018: 17 · 2017: 18 · 2016: 25 · 2015: 25 · 2014: 11 · 2013: 4
- FWCI
- 9.70
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Nucleic Acids Research (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- National Natural Science Foundation of China (31171247, 31271418)
- Fudan University (pNL4-3)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- zinc finger nucleases, HIV-1 latency, long terminal repeat, HIV cure, human T cells
- MeSH terms
- Cells, Cultured, DNA, Viral, Endodeoxyribonucleases, Humans, Proviruses, T-Lymphocytes, HIV-1, HIV Long Terminal Repeat, Zinc Fingers, Virus Integration, Sequence Deletion, Virus Latency, Jurkat Cells
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