Unknown,Transcriptomics,Genomics,Proteomics

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UPRT Transgenic cells, Human Arrays


ABSTRACT: UPRT Transgenic and wildtype control cells of both HeLa and U373MG tissue culture lines were grown in the presence of 2mM 2,4-dithiouracil for either 6 hours (HeLa) or 8 hours (U373MG) and total RNA, not thiouracil-purified, from both transgenic and wildtype samples was compared on microarrays. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Computed

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Cleary 

PROVIDER: E-GEOD-2948 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Biosynthetic labeling of RNA with uracil phosphoribosyltransferase allows cell-specific microarray analysis of mRNA synthesis and decay.

Cleary Michael D MD   Meiering Christopher D CD   Jan Eric E   Guymon Rebecca R   Boothroyd John C JC  

Nature biotechnology 20050130 2


Standard microarrays measure mRNA abundance, not mRNA synthesis, and therefore cannot identify the mechanisms that regulate gene expression. We have developed a method to overcome this limitation by using the salvage enzyme uracil phosphoribosyltransferase (UPRT) from the protozoan Toxoplasma gondii. T. gondii UPRT has been well characterized because of its application in monitoring parasite growth: mammals lack this enzyme activity and thus only the parasite incorporates (3)H-uracil into its nu  ...[more]

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