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  Screening of human gene promoter activities using transfected-cell arrays

Cheng, X., Guerasimova, A., Manke, T., Rosenstiel, P., Haas, S., Warnatz, H. J., et al. (2010). Screening of human gene promoter activities using transfected-cell arrays. Gene, 450(1-2), 48-54. doi:10.1016/j.gene.2009.10.003.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7C18-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7C19-9
Genre: Journal Article

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 Creators:
Cheng, X.1, Author
Guerasimova, A.2, Author              
Manke, T.3, Author              
Rosenstiel, P., Author
Haas, S.4, Author              
Warnatz, H. J.5, Author              
Querfurth, R.3, Author              
Nietfeld, W.2, Author              
Vanhecke, D.1, Author
Lehrach, H.2, Author              
Yaspo, M. L.5, Author              
Janitz, M.2, Author              
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              
3Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433547              
4Gene Structure and Array Design (Stefan Haas), Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479640              
5Human Chromosome 21 (Marie-Laure Yaspo), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479652              

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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
 Abstract: 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.

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Language(s): eng - English
 Dates: 2010-01-15
 Publication Status: Published in print
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Title: Gene
Source Genre: Journal
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Pages: - Volume / Issue: 450 (1-2) Sequence Number: - Start / End Page: 48 - 54 Identifier: ISSN: 1879-0038 (Electronic) 0378-1119 (Linking) %R S0378-1119(09)00543-5 [pii] 10.1016/j.gene.2009.10.003