Academic Journal
Comparative AAV-eGFP Transgene Expression Using Vector Serotypes 1–9, 7m8, and 8b in Human Pluripotent Stem Cells, RPEs, and Human and Rat Cortical Neurons
| العنوان: | Comparative AAV-eGFP Transgene Expression Using Vector Serotypes 1–9, 7m8, and 8b in Human Pluripotent Stem Cells, RPEs, and Human and Rat Cortical Neurons |
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| المؤلفون: | Thu T. Duong, James Lim, Vidyullatha Vasireddy, Tyler Papp, Hung Nguyen, Lanfranco Leo, Jieyan Pan, Shangzhen Zhou, H. Isaac Chen, Jean Bennett, Jason A. Mills |
| المصدر: | Stem Cells International, Vol 2019 (2019) |
| بيانات النشر: | Hindawi Limited, 2019. |
| سنة النشر: | 2019 |
| المجموعة: | LCC:Internal medicine |
| مصطلحات موضوعية: | Internal medicine, RC31-1245 |
| الوصف: | Recombinant adeno-associated virus (rAAV), produced from a nonpathogenic parvovirus, has become an increasing popular vector for gene therapy applications in human clinical trials. However, transduction and transgene expression of rAAVs can differ across in vitro and ex vivo cellular transduction strategies. This study compared 11 rAAV serotypes, carrying one reporter transgene cassette containing a cytomegalovirus immediate-early enhancer (eCMV) and chicken beta actin (CBA) promoter driving the expression of an enhanced green-fluorescent protein (eGFP) gene, which was transduced into four different cell types: human iPSC, iPSC-derived RPE, iPSC-derived cortical, and dissociated embryonic day 18 rat cortical neurons. Each cell type was exposed to three multiplicity of infections (MOI: 1E4, 1E5, and 1E6 vg/cell). After 24, 48, 72, and 96 h posttransduction, GFP-expressing cells were examined and compared across dosage, time, and cell type. Retinal pigmented epithelium showed highest AAV-eGFP expression and iPSC cortical the lowest. At an MOI of 1E6 vg/cell, all serotypes show measurable levels of AAV-eGFP expression; moreover, AAV7m8 and AAV6 perform best across MOI and cell type. We conclude that serotype tropism is not only capsid dependent but also cell type plays a significant role in transgene expression dynamics. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1687-966X 1687-9678 |
| Relation: | https://doaj.org/toc/1687-966X; https://doaj.org/toc/1687-9678 |
| DOI: | 10.1155/2019/7281912 |
| URL الوصول: | https://doaj.org/article/81d0b03f3f4247b2a10601923f3480b4 |
| رقم الانضمام: | edsdoj.81d0b03f3f4247b2a10601923f3480b4 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 1687966X 16879678 |
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| DOI: | 10.1155/2019/7281912 |