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          Institute: MPI für molekulare Genetik     Collection: Department of Computational Molecular Biology     Display Documents



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ID: 539696.0, MPI für molekulare Genetik / Department of Computational Molecular Biology
Screening of human gene promoter activities using transfected-cell arrays
Authors:Cheng, X.; Guerasimova, A.; Manke, T.; Rosenstiel, P.; Haas, S.; Warnatz, H. J.; Querfurth, R.; Nietfeld, W.; Vanhecke, D.; Lehrach, H.; Yaspo, M. L.; Janitz, M.
Language:English
Date of Publication (YYYY-MM-DD):2010-01-15
Title of Journal:Gene
Volume:450
Issue / Number:1-2
Start Page:48
End Page:54
Copyright:© 2010 Elsevier B.V.
Review Status:not specified
Audience:Experts Only
Abstract / Description:Promoters are the best characterized transcriptional regulatory sequences in complex genomes because of their predictable location immediately upstream of transcription start sites. Despite a substantial body of literature describing transcriptional promoters, the identification of true start sites for all human transcripts is far from complete. The same is true of the key structural and functional elements responsible for promoter action in different cell types. In order to identify elements responsible for promoter activity, we applied transfected-cell array technology to functionally evaluate promoters for genes involved in inflammatory bowel disease. Seventy-four promoters were examined by reverse transfection of a promoter-fluorescent reporter constructs into a human embryonic kidney cell line (HEK293T). Sixteen (21.6%) promoters were found to be active in HEK293 T cells. Correlations between promoter activity and endogenous transcript level were calculated, and 75% of active promoters were found to be associated with transcriptional activity of their gene counterparts. These results provide experimental evidence of promoter activity, which may aid in understanding the regulation of gene expression. Moreover, this is the first large-scale functional study of regulatory sequences to use a high-throughput transfected-cell array technique.
Free Keywords:Base Sequence; Cell Line; Cloning, Molecular; Gene Expression Profiling/*methods; *Gene Expression Regulation; Humans; Promoter Regions, Genetic/*genetics; Tissue Array Analysis/*methods; Transcription, Genetic; Transfection
External Publication Status:published
Document Type:Article
Communicated by:Martin Vingron
Affiliations:MPI für molekulare Genetik
External Affiliations:Free University Berlin, Department of Biology, Chemistry and Pharmacy, Takustr. 3, 14195 Berlin, Germany
Institut for Clinical Molecular Biology, Christian-Albrechts-University, Schittenhelmstr. 12, 24105 Kiel, Germany
Identifiers:ISSN:1879-0038 (Electronic) 0378-1119 (Linking) %R S037... [ID No:1]
URL:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=... [ID No:2]
DOI:10.1016/j.gene.2009.10.003
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