Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes
The level of transduction efficiency of the target retinal cells affects the choice of AAV serotype and the outcome of gene replacement therapy for inherited retinal diseases. This study focused on the tropism and transduction efficiency of AAV2.7m8-, AAV5-, AAV8-, and AAV9-GFP in ARPE-19 and HEK293...
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MDPI AG
2024-01-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/16/1/138 |
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| author | Dzerassa Gurtsieva Ekaterina Minskaia Sofia Zhuravleva Elena Subcheva Elena Sakhibgaraeva Andrew Brovin Artem Tumaev Alexander Karabelsky |
| author_facet | Dzerassa Gurtsieva Ekaterina Minskaia Sofia Zhuravleva Elena Subcheva Elena Sakhibgaraeva Andrew Brovin Artem Tumaev Alexander Karabelsky |
| author_sort | Dzerassa Gurtsieva |
| collection | DOAJ |
| description | The level of transduction efficiency of the target retinal cells affects the choice of AAV serotype and the outcome of gene replacement therapy for inherited retinal diseases. This study focused on the tropism and transduction efficiency of AAV2.7m8-, AAV5-, AAV8-, and AAV9-GFP in ARPE-19 and HEK293 cells. Fluorescence intensity was assessed bi-hourly by means of IncuCyte S3 live imaging microscopy. Within 12 h, AAV2.7m8 demonstrated the highest transduction efficiency at four viral concentrations of 1-, 3-, 6-, and 8 × 10<sup>4</sup> VG/cell in a dose-dependent manner, followed by AAV5 in ARPE-19 and AAV9 in HEK293 cells. The transduction efficiency of AAV2.7m8 at a dose of 6 × 10<sup>4</sup> VG/cell was 21, 202, and 323 times higher in ARPE-19 cells and 324, 100, and 52 times higher in HEK293 cells compared to AAV5, AAV8, and AAV9, respectively. This trend remained for 4 days at all viral concentrations, as additionally shown by flow cytometry. At a dose of 6 × 10<sup>4</sup> VG/cell, AAV2.7m8 (97% GFP-positive cells, GFP +) was nearly two and 10 times as efficient as AAV5 (52% GFP+) and AAV9 or AAV8 (both 9%), respectively, in ARPE-19 cells. In HEK293 cells, 95% of AAV2.7m8-, 26% of AAV9-, 17% of AAV8-, and 12% of AAV5-transduced cells were GFP-positive. |
| first_indexed | 2024-03-08T10:37:08Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-08T10:37:08Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-1c42311fcf654b39b5d703fc771d02242024-01-26T18:08:21ZengMDPI AGPharmaceutics1999-49232024-01-0116113810.3390/pharmaceutics16010138Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 SerotypesDzerassa Gurtsieva0Ekaterina Minskaia1Sofia Zhuravleva2Elena Subcheva3Elena Sakhibgaraeva4Andrew Brovin5Artem Tumaev6Alexander Karabelsky7Department of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaDepartment of Gene Therapy, Sirius University of Science and Technology, Olympic Avenue, 1, 354340 Sochi, RussiaThe level of transduction efficiency of the target retinal cells affects the choice of AAV serotype and the outcome of gene replacement therapy for inherited retinal diseases. This study focused on the tropism and transduction efficiency of AAV2.7m8-, AAV5-, AAV8-, and AAV9-GFP in ARPE-19 and HEK293 cells. Fluorescence intensity was assessed bi-hourly by means of IncuCyte S3 live imaging microscopy. Within 12 h, AAV2.7m8 demonstrated the highest transduction efficiency at four viral concentrations of 1-, 3-, 6-, and 8 × 10<sup>4</sup> VG/cell in a dose-dependent manner, followed by AAV5 in ARPE-19 and AAV9 in HEK293 cells. The transduction efficiency of AAV2.7m8 at a dose of 6 × 10<sup>4</sup> VG/cell was 21, 202, and 323 times higher in ARPE-19 cells and 324, 100, and 52 times higher in HEK293 cells compared to AAV5, AAV8, and AAV9, respectively. This trend remained for 4 days at all viral concentrations, as additionally shown by flow cytometry. At a dose of 6 × 10<sup>4</sup> VG/cell, AAV2.7m8 (97% GFP-positive cells, GFP +) was nearly two and 10 times as efficient as AAV5 (52% GFP+) and AAV9 or AAV8 (both 9%), respectively, in ARPE-19 cells. In HEK293 cells, 95% of AAV2.7m8-, 26% of AAV9-, 17% of AAV8-, and 12% of AAV5-transduced cells were GFP-positive.https://www.mdpi.com/1999-4923/16/1/138IRDretinopathygene therapyAAV serotypesAAV tropismretina |
| spellingShingle | Dzerassa Gurtsieva Ekaterina Minskaia Sofia Zhuravleva Elena Subcheva Elena Sakhibgaraeva Andrew Brovin Artem Tumaev Alexander Karabelsky Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes Pharmaceutics IRD retinopathy gene therapy AAV serotypes AAV tropism retina |
| title | Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes |
| title_full | Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes |
| title_fullStr | Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes |
| title_full_unstemmed | Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes |
| title_short | Engineered AAV2.7m8 Serotype Shows Significantly Higher Transduction Efficiency of ARPE-19 and HEK293 Cell Lines Compared to AAV5, AAV8 and AAV9 Serotypes |
| title_sort | engineered aav2 7m8 serotype shows significantly higher transduction efficiency of arpe 19 and hek293 cell lines compared to aav5 aav8 and aav9 serotypes |
| topic | IRD retinopathy gene therapy AAV serotypes AAV tropism retina |
| url | https://www.mdpi.com/1999-4923/16/1/138 |
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