Screening for novel human genes associated with CRE pathway activation with cell microarray.
In this study, cell microarray technology is used to identify novel human genes associated with CRE pathway activation. By reverse transfection, expression plasmids containing full-length cDNAs were cotransfected with the reporter plasmid pCRE-d2EGFP to monitor the activation of the CRE pathway via enhanced green fluorescence protein (EGFP) expression. Of the 575 predominantly novel genes screened, 22 exhibited relatively higher EGFP fluorescence compared with a negative control. After a functional validation with a dual luciferase reporter system that included both cis- and trans-luciferase assays, 4 of the 22 genes (RNF41, C8orf32, C6orf208, and MEIS3P1) were confirmed as CRE-pathway activators. Western blot analysis revealed that RNF41 can promote CREB phosphorylation. These results demonstrate the successful combination of cell microarray technology with this reporting system and the potential of this tool to characterize functions of novel genes in a highly parallel format.
Tian
Linjie
L
Chinese National Human Genome Center, Beijing, 3-707 North YongChang Road BDA, Beijing 100176, People's Republic of China.
Wang
Pingzhang
P
Guo
Jinhai
J
Wang
Xinyu
X
Deng
Weiwei
W
Zhang
Chenying
C
Fu
Dongxu
D
Gao
Xia
X
Shi
Taiping
T
Ma
Dalong
D
D002460:Cell Line; D000242:Cyclic AMP / Q000235:genetics* / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D019426:Integrases / Q000235:genetics; D020218:Response Elements / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics*; D046888:Tissue Array Analysis
D000242:Cyclic AMP; C045073:Cre recombinase; D019426:Integrases