TY - JOUR
T1 - Rapid analysis of gene expression (RAGE) facilitates universal expression profiling
AU - Wang, Aijin
AU - Pierce, Angela
AU - Judson-Kremer, Kimberly
AU - Gaddis, Sara
AU - Aldaz, C. Marcelo
AU - Johnson, David G.
AU - MacLeod, Michael C.
N1 - Funding Information:
We gratefully acknowledge the help of Rebecca Deen, Michelle Gardiner, Judy Ing and Chris Yone in the preparation of this manuscript. This work was supported by grants from the American Cancer Society (RPG-96-001-03-CNE to M.C.M.) and the National Institutes of Health (CA79648 to D.G.J., CA35581 to M.C.M.), and by NIEHS Center Grant ES07784 and NCI Cancer Center Grant CA16672.
PY - 1999/12/1
Y1 - 1999/12/1
N2 - Current techniques for analysis of gene expression either monitor one gene at a time, for example northern hybridization or RT-PCR methods, or are designed for the simultaneous analysis of thousands of genes, for example microarray hybridization or serial analysis of gene expression. To provide a flexible, intermediate scale alternative, a PCR-based method for the rapid analysis of gene expression has been developed which allows expression changes to be determined in either a directed search of known genes, or an undirected survey of unknown genes. A single set of reagents and reaction conditions allows analyses of most genes in any eukaryote. The method is useful for assaying on the order of tens to hundreds of genes in multiple samples. Control experiments indicate reliable detection of changes in gene expression 2-fold and greater, and sensitivity of detection better than 1 in 10,000. Analyses of over 400 genes in a mouse system transgenic for the E2F1 gene have identified several new downstream targets of E2F1, including Brca1 and Cdk7, in addition to several unidentified genes that are upregulated in the transgenic mice. Changes in expression of several genes related to apoptosis suggest a possible potentiation of apoptotic pathways in the transgenic keratinocytes.
AB - Current techniques for analysis of gene expression either monitor one gene at a time, for example northern hybridization or RT-PCR methods, or are designed for the simultaneous analysis of thousands of genes, for example microarray hybridization or serial analysis of gene expression. To provide a flexible, intermediate scale alternative, a PCR-based method for the rapid analysis of gene expression has been developed which allows expression changes to be determined in either a directed search of known genes, or an undirected survey of unknown genes. A single set of reagents and reaction conditions allows analyses of most genes in any eukaryote. The method is useful for assaying on the order of tens to hundreds of genes in multiple samples. Control experiments indicate reliable detection of changes in gene expression 2-fold and greater, and sensitivity of detection better than 1 in 10,000. Analyses of over 400 genes in a mouse system transgenic for the E2F1 gene have identified several new downstream targets of E2F1, including Brca1 and Cdk7, in addition to several unidentified genes that are upregulated in the transgenic mice. Changes in expression of several genes related to apoptosis suggest a possible potentiation of apoptotic pathways in the transgenic keratinocytes.
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U2 - 10.1093/nar/27.23.4609
DO - 10.1093/nar/27.23.4609
M3 - Article
C2 - 10556317
AN - SCOPUS:0033485487
SN - 0305-1048
VL - 27
SP - 4609
EP - 4618
JO - Nucleic acids research
JF - Nucleic acids research
IS - 23
ER -