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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
2006-4-27
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pubmed:abstractText |
A primary limitation in the development and use of screens to identify factors that regulate mammalian pre-mRNA splicing has been the development of sensitive reporter assays. Alternative splicing typically involves relatively small (< 10-fold) changes in isoform ratios. Therefore, reporter constructs designed to allow direct analysis of isoform expression historically have at most a 10-fold window of discrimination between a positive signal and background. Here we describe the design and application of a reporter cell line that makes use of the phenomenon of transcriptional synergy to amplify the detection of changes in splicing, such that a three- to five-fold change in splicing pattern is observed as a 30- to 50-fold change in GFP expression. Using this cell line we have identified two small molecules, from a library of approximately 300 synthetic compounds, that can induce partial repression of a variable exon from the CD45 gene. We propose that the concept of transcription-based amplification of signal will allow the development of true high-throughput screening approaches to identify effectors of mammalian alternative splicing.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-10594010,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-11483522,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-12515380,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-12526676,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-12626338,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-14636588,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-14684825,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-14705182,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-1511877,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-15777182,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-16156783,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-2160609,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-3047590,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16556940-8947053
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
1355-8382
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
12
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
925-30
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:16556940-Alternative Splicing,
pubmed-meshheading:16556940-Antigens, CD45,
pubmed-meshheading:16556940-Cell Line,
pubmed-meshheading:16556940-Exons,
pubmed-meshheading:16556940-Gene Library,
pubmed-meshheading:16556940-Genes, Reporter,
pubmed-meshheading:16556940-Green Fluorescent Proteins,
pubmed-meshheading:16556940-Luminescent Proteins,
pubmed-meshheading:16556940-Lymphocyte Activation,
pubmed-meshheading:16556940-RNA Precursors,
pubmed-meshheading:16556940-RNA Splicing,
pubmed-meshheading:16556940-Sensitivity and Specificity,
pubmed-meshheading:16556940-Signal Transduction,
pubmed-meshheading:16556940-T-Lymphocytes,
pubmed-meshheading:16556940-Tetradecanoylphorbol Acetate,
pubmed-meshheading:16556940-Transcription, Genetic,
pubmed-meshheading:16556940-Transfection
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pubmed:year |
2006
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pubmed:articleTitle |
Use of transcriptional synergy to augment sensitivity of a splicing reporter assay.
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pubmed:affiliation |
Department of Biochemistry, UT Southwestern Medical Center, Dallas, Texas 75390-9038, USA
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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