Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo.

Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo. Research Abstract Details 

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Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo. Abstract Text:

Mohammed Al-Dosari,Guisheng Zhang,Joseph E Knapp,Dexi Liu,

Although transcription regulation of human cytochrome P450 enzymes (CYP) is known to play an important role in drug metabolism and homeostasis, factors influencing the expression of various CYP genes in humans remain largely undefined. We used three cell lines and CD-1 mice to assess the activity of genomic promoter sequences of human CYP2D6, 1A2, 3A4, 2C9, 2C18, and 2E1 genes. CYP promoter sequences were amplified by PCR using human liver genomic DNA as the template and cloned into pGL3-Basic vectors that contain a luciferase reporter gene but lack promoter or enhancer sequences. Each plasmid construct was transfected into cells in vitro using polyethylenimine (PEI) as the transfection reagent and into mice using the recently developed hydrodynamics-based procedure. Relative promoter strength was determined by the level of luciferase expression in transfected cells. All six human CYP promoters are active in driving reporter gene expression in cultured hepatic HepG2 cells and non-hepatic cells such as human embryonic kidney fibroblasts (293 cells) and murine melanoma cells (BL-16) as well as cells in intact mouse liver, lung, heart, kidney and spleen. The order of strength among CYP promoters examined was found to be 2D6 > 1A2 > 3A4 > 2C9 > 2C18 > 2E1.

Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo. Publishing Authors By Initials

M Al-Dosari,G Zhang,JE Knapp,D Liu,

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Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Bioscience reports

VOLUME: 26

Page Numbers: 217-29

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ISSN: 0144-8463

DAY: 3

MONTH: Jun

YEAR: 2006

Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo. Information

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LANGUAGE: eng

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Grant and Affiliation Information for Direct assessment of promoter activity of human cytochrome p450 genes using optimized transfection in vitro and in vivo.

AFFILIATION: Department of Pharmaceutical Sciences, University of Pittsburgh School of Pharmacy, 527 Salk Hall, Pittsburgh, PA 15261, USA.

Country: United States

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MEDLINETA: Biosci Rep

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