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 develo...
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| Format: | Article |
| Language: | English |
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BMC
2020-07-01
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| Series: | Cell & Bioscience |
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| Online Access: | http://link.springer.com/article/10.1186/s13578-020-00444-w |
| _version_ | 1818369740776669184 |
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| author | Zhepeng Liu Jin Liang Shuliang Chen Kewu Wang Xianhao Liu Beibei Liu Yang Xia Mingxiong Guo Xiaoshi Zhang Guihong Sun Geng Tian |
| author_facet | Zhepeng Liu Jin Liang Shuliang Chen Kewu Wang Xianhao Liu Beibei Liu Yang Xia Mingxiong Guo Xiaoshi Zhang Guihong Sun Geng Tian |
| author_sort | Zhepeng Liu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T23:28:39Z |
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| id | doaj.art-08f0604161ac4926b9b74765cc2c7a3c |
| institution | Directory Open Access Journal |
| issn | 2045-3701 |
| language | English |
| last_indexed | 2024-12-13T23:28:39Z |
| publishDate | 2020-07-01 |
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| series | Cell & Bioscience |
| spelling | doaj.art-08f0604161ac4926b9b74765cc2c7a3c2022-12-21T23:27:28ZengBMCCell & Bioscience2045-37012020-07-0110111310.1186/s13578-020-00444-wGenome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infectionZhepeng Liu0Jin Liang1Shuliang Chen2Kewu Wang3Xianhao Liu4Beibei Liu5Yang Xia6Mingxiong Guo7Xiaoshi Zhang8Guihong Sun9Geng Tian10Department of Biotherapy Research Center, Sun Yat-sen University Cancer CenterDepartment of Oncology, The First Affiliated Hospital of Shenzhen University, The Second People’s Hospital of ShenzhenSchool of Basic Medical Sciences, Wuhan UniversityDepartment of Oncology, The Second People’s Hospital of WuhuDepartment of Oncology, The First Affiliated Hospital of Shenzhen University, The Second People’s Hospital of ShenzhenDepartment of Oncology, The First Affiliated Hospital of Shenzhen University, The Second People’s Hospital of ShenzhenDepartment of Oncology, The First Affiliated Hospital of Shenzhen University, The Second People’s Hospital of ShenzhenCollege of Life Sciences, Wuhan UniversityDepartment of Biotherapy Research Center, Sun Yat-sen University Cancer CenterSchool of Basic Medical Sciences, Wuhan UniversityDepartment of Oncology, The First Affiliated Hospital of Shenzhen University, The Second People’s Hospital of ShenzhenAbstract 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.http://link.springer.com/article/10.1186/s13578-020-00444-wCCR5HIV-1CRISPR/AsCpf1HIV-1 resistanceSelective advantage |
| spellingShingle | Zhepeng Liu Jin Liang Shuliang Chen Kewu Wang Xianhao Liu Beibei Liu Yang Xia Mingxiong Guo Xiaoshi Zhang Guihong Sun Geng Tian Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection Cell & Bioscience CCR5 HIV-1 CRISPR/AsCpf1 HIV-1 resistance Selective advantage |
| title | Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection |
| title_full | Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection |
| title_fullStr | Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection |
| title_full_unstemmed | Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection |
| title_short | Genome editing of CCR5 by AsCpf1 renders CD4+T cells resistance to HIV-1 infection |
| title_sort | genome editing of ccr5 by ascpf1 renders cd4 t cells resistance to hiv 1 infection |
| topic | CCR5 HIV-1 CRISPR/AsCpf1 HIV-1 resistance Selective advantage |
| url | http://link.springer.com/article/10.1186/s13578-020-00444-w |
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