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An enhancer/promoter combination strengthens the expression of blood-coagulation factor VIII in non-viral expression vectors
Picanço-Castro, V; Russo-Carbolante, E. M. S; Fontes, A. M; Fernandes, A. C; Covas, D. T.
  • Picanço-Castro, V; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Hemocentro de Ribeirão Preto. Ribeirão Preto. BR
  • Russo-Carbolante, E. M. S; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Hemocentro de Ribeirão Preto. Ribeirão Preto. BR
  • Fontes, A. M; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Hemocentro de Ribeirão Preto. Ribeirão Preto. BR
  • Fernandes, A. C; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Hemocentro de Ribeirão Preto. Ribeirão Preto. BR
  • Covas, D. T; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Hemocentro de Ribeirão Preto. Ribeirão Preto. BR
Genet. mol. res. (Online) ; 7(2): 314-325, 2008. tab, ilus
Article in English | LILACS | ID: lil-641008
ABSTRACT
We explored the potential of fusion of hepatic locus control region 1 (HCR-1) with HCR-2 to express B-domain-deleted human factor VIII (FVIII) in four cell lines. B-domain-deleted human FVIII expression was controlled by HCR-1/HCR-2, followed by liver specific and ubiquitous promoters. Chimera enhancer HCR-1/HCR-2, followed by cytomegalovirus (CMV) promoter, gave 2-fold more FVIII expression in all cell lines (105.6 ± 2.8 for Hek-293, 68.8 ± 3.8 for HepG2, 34.8 ± 1.3 for CHO, and 27.2 ± 1.6 ng-mL-1-106 cells-1 for L.N.) when compared to the vector with CMV alone (54.8 ± 3.3 for Hek-293, 32.4 ± 1.2 for HepG2, 18.6 ± 1.1 for CHO, and 10.1 ± 1.7 ng-mL-1-106 cells-1 for L.N.). Elongation factor 1gene and human CMV promoters were more efficient than the promoters from the human α-1-antitrypsin gene, and fviii was less efficient in hepatic cell lines. HCR-1/HCR-2, followed by strong promoters, increases FVIII expression in vitro. Our results underscore the importance of cis sequences for enhancing in vitro FVIII expression; this may be helpful for designing new strategies to improve heterologous expression systems.
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Full text: Available Index: LILACS (Americas) Main subject: Factor VIII / Enhancer Elements, Genetic / Promoter Regions, Genetic / Genetic Vectors Limits: Animals / Humans Language: English Journal: Genet. mol. res. (Online) Journal subject: Molecular Biology / Genetics Year: 2008 Type: Article Affiliation country: Brazil Institution/Affiliation country: Universidade de São Paulo/BR

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Full text: Available Index: LILACS (Americas) Main subject: Factor VIII / Enhancer Elements, Genetic / Promoter Regions, Genetic / Genetic Vectors Limits: Animals / Humans Language: English Journal: Genet. mol. res. (Online) Journal subject: Molecular Biology / Genetics Year: 2008 Type: Article Affiliation country: Brazil Institution/Affiliation country: Universidade de São Paulo/BR