Academic Journal
Factor VIII mRNA expression from a BAC carrying the intact locus made by homologous recombination
| العنوان: | Factor VIII mRNA expression from a BAC carrying the intact locus made by homologous recombination |
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| المؤلفون: | Pérez Luz, Sara, Abdulrazzak, H, Grillot-Courvalin, C, Huxley, C |
| سنة النشر: | 2007 |
| المجموعة: | Universidad Complutense de Madrid (UCM): E-Prints Complutense |
| مصطلحات موضوعية: | 636.09, Hemophilia A, Factor VIII, F8, Gene therapy, Bacterial artificial chromosome, BAC, Bacterial delivery, OriP/EBNA-1, Alphoid DNA, Biología, Veterinaria, 2415 Biología Molecular, 3109 Ciencias Veterinarias |
| Time: | 577 |
| الوصف: | Hemophilia A is caused by mutations in the gene encoding factor VIII (F8) and is an important target for gene therapy. The F8 gene contains 26 exons spread over ∼186 kb and no work using the intact genomic locus has been carried out. We have constructed a 250-kb BAC carrying all 26 exons, the introns, and more than 40 kb of upstream and 20 kb of downstream DNA. This F8 BAC was further retrofitted with either the oriP/EBNA-1 elements from Epstein–Barr virus, which allow episomal maintenance in mammalian cells, or alphoid DNA, which allows human artificial chromosome formation in some human cell lines. Lipofection of the oriP/EBNA-1-containing version into mouse Hepa1-6 cells resulted in expression of F8 mRNA spanning the F8 gene. The >300-kb BAC carrying alphoid DNA was successfully delivered to 293A and HT1080 cells using bacterial delivery, resulting in greater than endogenous levels of F8 mRNA expression. ; Ministerio de Educación y Ciencia de España (EX2003-0291) ; Medical Research Council, United Kingdom ; EU Training Network HPRN CT2000-00089 (MAKEMAC) ; Depto. de Bioquímica y Biología Molecular ; Fac. de Veterinaria ; TRUE ; pub |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 0888-7543 |
| Relation: | https://hdl.handle.net/20.500.14352/93001; https://www.sciencedirect.com/science/article/pii/S0888754307001759?via%3Dihub |
| DOI: | 10.1016/j.ygeno.2007.07.005 |
| الاتاحة: | https://hdl.handle.net/20.500.14352/93001 https://doi.org/10.1016/j.ygeno.2007.07.005 https://www.sciencedirect.com/science/article/pii/S0888754307001759?via%3Dihub |
| Rights: | Attribution 4.0 International ; http://creativecommons.org/licenses/by/4.0/ ; open access |
| رقم الانضمام: | edsbas.EB547DAA |
| قاعدة البيانات: | BASE |
Full Text Finder | تدمد: | 08887543 |
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| DOI: | 10.1016/j.ygeno.2007.07.005 |