title: Screening for novel human genes associated with CRE pathway activation with cell microarray.
abstract: 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.
authors.affiliation: Chinese National Human Genome Center, Beijing, 3-707 North YongChang Road BDA, Beijing 100176, People's Republic of China.
authors.forename: Linjie
authors.initials: L
authors.lastname: Tian
authors.forename: Pingzhang
authors.initials: P
authors.lastname: Wang
authors.forename: Jinhai
authors.initials: J
authors.lastname: Guo
authors.forename: Xinyu
authors.initials: X
authors.lastname: Wang
authors.forename: Weiwei
authors.initials: W
authors.lastname: Deng
authors.forename: Chenying
authors.initials: C
authors.lastname: Zhang
authors.forename: Dongxu
authors.initials: D
authors.lastname: Fu
authors.forename: Xia
authors.initials: X
authors.lastname: Gao
authors.forename: Taiping
authors.initials: T
authors.lastname: Shi
authors.forename: Dalong
authors.initials: D
authors.lastname: Ma
mesh_terms: 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
chemical_list: D000242:Cyclic AMP; C045073:Cre recombinase; D019426:Integrases