Disruption of HIV-1 co-receptors CCR5 and CXCR4 in primary human T cells and hematopoietic stem and progenitor cells using base editing
| Autor: | Alejandra Gutierrez-Guerrero, Cindy R. Eide, Jakub Tolar, David R. Liu, Kyle D. Smith, Benjamin Steinbeck, Friederike Knipping, Gregory A. Newby, Mark J. Osborn, Els Verhoeyen, Colby J. Feser, Yongxing Fang, Claudio Mussolino, Caroline Costa, Bruce R. Blazar, Anton P. McCaffrey, Keli L. Hippen, Amber N. McElroy, Samuel P. Bingea, Sarah C. Nielsen, Tatjana I. Cornu |
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| Rok vydání: | 2021 |
| Předmět: |
Pharmacology
Gene Editing Receptors CXCR4 Receptors CCR5 Point mutation T-Lymphocytes HIV Infections Biology Hematopoietic Stem Cells CXCR4 Stop codon Viral vector Cell biology Transduction (genetics) Start codon Drug Discovery Genetics HIV-1 Molecular Medicine Humans Original Article Progenitor cell Molecular Biology Gene |
| Zdroj: | Mol Ther |
| ISSN: | 1525-0024 |
| Popis: | Disruption of CCR5 or CXCR4, the main human immunodeficiency virus type 1 (HIV-1) co-receptors, has been shown to protect primary human CD4(+) T cells from HIV-1 infection. Base editing can install targeted point mutations in cellular genomes, and can thus efficiently inactivate genes by introducing stop codons or eliminating start codons without double-stranded DNA break formation. Here, we applied base editors for individual and simultaneous disruption of both co-receptors in primary human CD4(+) T cells. Using cytosine base editors we observed premature stop codon introduction in up to 89% of sequenced CCR5 or CXCR4 alleles. Using adenine base editors we eliminated the start codon in CCR5 in up to 95% of primary human CD4(+) T cell and up to 88% of CD34(+) hematopoietic stem and progenitor cell target alleles. Genome-wide specificity analysis revealed low numbers of off-target mutations that were introduced by base editing, located predominantly in intergenic or intronic regions. We show that our editing strategies prevent transduction with CCR5-tropic and CXCR4-tropic viral vectors in up to 79% and 88% of human CD4(+) T cells, respectively. The engineered T cells maintained functionality and overall our results demonstrate the effectiveness of base-editing strategies for efficient and specific ablation of HIV co-receptors in clinically relevant cell types. |
| Databáze: | OpenAIRE |
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