Identification of five human novel genes associated with cell proliferation by cell-based screening from an expressed cDNA ORF library

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• 作者：Xi Ma ; Xinyu Wang ; Xia Gao ; Lan Wang ; Yang Lu ; Peng Gao ; Weiwei Deng ; Peng Yu ; Jinjing Ma ; Jinhai Guo ; Hualing Cheng ; Chenying Zhang ; Taiping Shi ; Dalong Ma
• 关键词：High-throughput screening ; Cell proliferation ; Reporter gene ; Functional genomics
• 刊名：Life Sciences
• 出版年：2007
• 出版时间：15 September 2007
• 年：2007
• 卷：81
• 期：14
• 页码：1141-1151
• 全文大小：1551 K

文摘

The development of functional profiling technologies provides opportunity for high-throughput functional genomics studies. We describe a cell-based screening system to identify novel human genes associated with cell proliferation. The method integrates luciferase reporter gene activity, fluorescence stain, automated microscopy and cellular phenotype assays. We successfully used the system to screen 409 novel human genes cloned by our lab and found that 27 genes significantly up-regulated promoter-Renilla luciferase reporter plasmid (pRL) activity. Among them, five genes, TRAF3IP3, ZNF306, ZNF250, SGOL1, and ZNF434, were determined through morphological observation, calcein AM fluorescence stain, MTT assay and cell cycle analysis to be associated with cell proliferation. Furthermore, we showed that the gene TRAF3IP3, which initially was identified to specifically interact with TRAF3, stimulated cell growth by modulating the c-Jun N-terminal kinase (JNK) pathway, and RNAi of TRAF3IP3 confirmed that the effect was physiological and necessary. In summary, we integrated a rapid and efficient system for screening novel growth regulatory genes. Using the new screening system we identified five genes associated with cell proliferation for the first time.