Dataset Information

Genome editing of CCR5 by AsCpf1 renders CD4⁺T cells resistance to HIV-1 infection.

ABSTRACT:

Background

The chemokine receptor CCR5 is one of the co-receptor of HIV-1 infection. People with homozygous CCR5Δ32 deletion resist HIV-1 infection, which makes the CCR5 an important target for HIV-1 gene therapy. Although the CRISPR/Cas9 has ever been used for HIV-1 study, the newly developed CRISPR/AsCpf1 has never been utilized in HIV-1 co-receptor disruption. The CRISPR/Cpf1 system shows many advantages over CRISPR/Cas9, such as lower off-target, small size of nuclease, easy sgRNA design for multiplex gene editing, etc. Therefore, the CRISPR/Cpf1 mediated gene editing will confer a more specific and safe strategy in HIV-1 co-receptor disruption.

Results

Here, we demonstrated that CRISPR/AsCpf1 could ablate the main co-receptor of HIV-1 infection-CCR5 efficiently with two screened sgRNAs via different delivery strategies (lentivirus, adenovirus). The edited cells resisted R5-tropic HIV-1 infection but not X4-tropic HIV-1 infection compared with the control group in different cell types of HIV-1 study (TZM.bl, SupT1-R5, Primary CD4⁺T cells). Meanwhile, the edited cells exhibited selective advantage over unedited cells while under the pressure of R5-tropic HIV-1. Furthermore, we clarified that the predicted off-target sites of selected sgRNAs were very limited, which is much less than regular using sgRNAs for CRISPR/Cas9, and no evident off-target was observed. We also showed that the disruption of CCR5 by CRISPR/AsCpf1 took no effects on cell proliferation and apoptosis.

Conclusions

Our study provides a basis for a possible application of CCR5-targeting gene editing by CRISPR/AsCpf1 with high specific sgRNAs against HIV-1 infection.

SUBMITTER: Liu Z

PROVIDER: S-EPMC7346486 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Publications

Genome editing of CCR5 by AsCpf1 renders CD4<sup>+</sup>T cells resistance to HIV-1 infection.

Liu Zhepeng Z Liang Jin J Chen Shuliang S Wang Kewu K Liu Xianhao X Liu Beibei B Xia Yang Y Guo Mingxiong M Zhang Xiaoshi X Sun Guihong G Tian Geng G

Cell & bioscience 20200708

<h4>Background</h4>The chemokine receptor CCR5 is one of the co-receptor of HIV-1 infection. People with homozygous <i>CCR5Δ32</i> deletion resist HIV-1 infection, which makes the <i>CCR5</i> an important target for HIV-1 gene therapy. Although the CRISPR/Cas9 has ever been used for HIV-1 study, the newly developed CRISPR/AsCpf1 has never been utilized in HIV-1 co-receptor disruption. The CRISPR/Cpf1 system shows many advantages over CRISPR/Cas9, such as lower off-target, small size of nuclease, ...[more]

PMID: 32670545

Similar Datasets

Project description:BackgroundCCR5 is the major coreceptor for human immunodeficiency virus (HIV). We investigated whether site-specific modification of the gene ("gene editing")--in this case, the infusion of autologous CD4 T cells in which the CCR5 gene was rendered permanently dysfunctional by a zinc-finger nuclease (ZFN)--is safe.MethodsWe enrolled 12 patients in an open-label, nonrandomized, uncontrolled study of a single dose of ZFN-modified autologous CD4 T cells. The patients had chronic aviremic HIV infection while they were receiving highly active antiretroviral therapy. Six of them underwent an interruption in antiretroviral treatment 4 weeks after the infusion of 10 billion autologous CD4 T cells, 11 to 28% of which were genetically modified with the ZFN. The primary outcome was safety as assessed by treatment-related adverse events. Secondary outcomes included measures of immune reconstitution and HIV resistance.ResultsOne serious adverse event was associated with infusion of the ZFN-modified autologous CD4 T cells and was attributed to a transfusion reaction. The median CD4 T-cell count was 1517 per cubic millimeter at week 1, a significant increase from the preinfusion count of 448 per cubic millimeter (P<0.001). The median concentration of CCR5-modified CD4 T cells at 1 week was 250 cells per cubic millimeter. This constituted 8.8% of circulating peripheral-blood mononuclear cells and 13.9% of circulating CD4 T cells. Modified cells had an estimated mean half-life of 48 weeks. During treatment interruption and the resultant viremia, the decline in circulating CCR5-modified cells (-1.81 cells per day) was significantly less than the decline in unmodified cells (-7.25 cells per day) (P=0.02). HIV RNA became undetectable in one of four patients who could be evaluated. The blood level of HIV DNA decreased in most patients.ConclusionsCCR5-modified autologous CD4 T-cell infusions are safe within the limits of this study. (Funded by the National Institute of Allergy and Infectious Diseases and others; ClinicalTrials.gov number, NCT00842634.).

Dataset Information

Genome editing of CCR5 by AsCpf1 renders CD4⁺T cells resistance to HIV-1 infection.

Background

Results

Conclusions

Publications

Genome editing of CCR5 by AsCpf1 renders CD4<sup>+</sup>T cells resistance to HIV-1 infection.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection.

Background

Results

Conclusions

Publications

Genome editing of CCR5 by AsCpf1 renders CD4<sup>+</sup>T cells resistance to HIV-1 infection.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Genome editing of CCR5 by AsCpf1 renders CD4⁺T cells resistance to HIV-1 infection.