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Genet. mol. res. (Online) ; Genet. mol. res. (Online);7(2): 314-325, 2008. tab, ilus
Artigo em Inglês | LILACS | ID: lil-641008

RESUMO

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 1-α gene 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.


Assuntos
Humanos , Animais , Elementos Facilitadores Genéticos/genética , Fator VIII/genética , Regiões Promotoras Genéticas/genética , Vetores Genéticos/genética , Linhagem Celular , Linhagem Celular Tumoral , Células CHO , Cricetinae , Cricetulus , Citomegalovirus/genética , Fator VIII/metabolismo , Imuno-Histoquímica , Microscopia de Fluorescência , Plasmídeos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
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