Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology
C-C chemokine receptor type 5 (CCR5) is the main co-receptor for HIV entry into the target CD4+ cells, and homozygous CCR5Δ32/Δ32 cells are resistant to CCR5-tropic HIV infection. However, the CCR5Δ32/Δ32 homozygous donors in populations are rare. Here we developed a simple approach to induce CCR5Δ3...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118301094 |
| _version_ | 1828849258365714432 |
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| author | Chunxia Qi Dan Li Xiangxiang Jiang Xiaopeng Jia Lingling Lu Yanfeng Wang Jinhuan Sun Yiming Shao Min Wei |
| author_facet | Chunxia Qi Dan Li Xiangxiang Jiang Xiaopeng Jia Lingling Lu Yanfeng Wang Jinhuan Sun Yiming Shao Min Wei |
| author_sort | Chunxia Qi |
| collection | DOAJ |
| description | C-C chemokine receptor type 5 (CCR5) is the main co-receptor for HIV entry into the target CD4+ cells, and homozygous CCR5Δ32/Δ32 cells are resistant to CCR5-tropic HIV infection. However, the CCR5Δ32/Δ32 homozygous donors in populations are rare. Here we developed a simple approach to induce CCR5Δ32/Δ32 homozygotes through CRISPR-Cas9 genome-editing technology. Designing a pair of single-guide RNA targeting the flank region of the CCR5Δ32 mutation locus, we applied the CRISPR-Cas9 and lentiviral packaging system to successfully convert wild-type CCR5 into CCR5Δ32/Δ32 homozygotes in the human Jurkat CD4+ cell line and primary CD4+ cells, exactly the same as the naturally occurring CCR5Δ32/Δ32 mutation. The successful rate is up to 20% in Jurkat cells but less in primary CD4+ cells. The modified CCR5Δ32/Δ32 CD4+ cells are resistant to CCR5-tropic HIV infection. Whole-genome sequencing revealed no apparent off-target sites. This approach has the promise to promote HIV/AIDS therapy from the only cured unique Berlin patient to a routine autologous cell-based therapy. Keywords: HIV, AIDS, CCR5Δ, 32, CRISPR-Cas9, CD4+ cells |
| first_indexed | 2024-12-12T22:47:11Z |
| format | Article |
| id | doaj.art-fbeed6f5c1314787bf56938ea071af03 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-12T22:47:11Z |
| publishDate | 2018-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-fbeed6f5c1314787bf56938ea071af032022-12-22T00:09:10ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-09-0112267274Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing TechnologyChunxia Qi0Dan Li1Xiangxiang Jiang2Xiaopeng Jia3Lingling Lu4Yanfeng Wang5Jinhuan Sun6Yiming Shao7Min Wei8School of Medicine, Nankai University, Tianjin, ChinaNational Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, ChinaSchool of Medicine, Nankai University, Tianjin, ChinaSchool of Medicine, Nankai University, Tianjin, ChinaSchool of Medicine, Nankai University, Tianjin, ChinaSchool of Medicine, Nankai University, Tianjin, ChinaSchool of Medicine, Nankai University, Tianjin, ChinaSchool of Medicine, Nankai University, Tianjin, China; National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, ChinaSchool of Medicine, Nankai University, Tianjin, China; Nankai University Second People's Hospital, School of Medicine, Nankai University, Tianjin, China; Corresponding author: Min Wei, Nankai University Second People’s Hospital, School of Medicine, Nankai University, No. 94 Weijin Road, Nankai District, Tianjin 300071, China.C-C chemokine receptor type 5 (CCR5) is the main co-receptor for HIV entry into the target CD4+ cells, and homozygous CCR5Δ32/Δ32 cells are resistant to CCR5-tropic HIV infection. However, the CCR5Δ32/Δ32 homozygous donors in populations are rare. Here we developed a simple approach to induce CCR5Δ32/Δ32 homozygotes through CRISPR-Cas9 genome-editing technology. Designing a pair of single-guide RNA targeting the flank region of the CCR5Δ32 mutation locus, we applied the CRISPR-Cas9 and lentiviral packaging system to successfully convert wild-type CCR5 into CCR5Δ32/Δ32 homozygotes in the human Jurkat CD4+ cell line and primary CD4+ cells, exactly the same as the naturally occurring CCR5Δ32/Δ32 mutation. The successful rate is up to 20% in Jurkat cells but less in primary CD4+ cells. The modified CCR5Δ32/Δ32 CD4+ cells are resistant to CCR5-tropic HIV infection. Whole-genome sequencing revealed no apparent off-target sites. This approach has the promise to promote HIV/AIDS therapy from the only cured unique Berlin patient to a routine autologous cell-based therapy. Keywords: HIV, AIDS, CCR5Δ, 32, CRISPR-Cas9, CD4+ cellshttp://www.sciencedirect.com/science/article/pii/S2162253118301094 |
| spellingShingle | Chunxia Qi Dan Li Xiangxiang Jiang Xiaopeng Jia Lingling Lu Yanfeng Wang Jinhuan Sun Yiming Shao Min Wei Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology Molecular Therapy: Nucleic Acids |
| title | Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology |
| title_full | Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology |
| title_fullStr | Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology |
| title_full_unstemmed | Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology |
| title_short | Inducing CCR5Δ32/Δ32 Homozygotes in the Human Jurkat CD4+ Cell Line and Primary CD4+ Cells by CRISPR-Cas9 Genome-Editing Technology |
| title_sort | inducing ccr5δ32 δ32 homozygotes in the human jurkat cd4 cell line and primary cd4 cells by crispr cas9 genome editing technology |
| url | http://www.sciencedirect.com/science/article/pii/S2162253118301094 |
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