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Cell-Based Screening and Validation of Human Novel Genes Associated with Cell Viability
1 Chinese National Human Genome Center, Beijing, China.; Lab of Medical Immunology, School of Basic Medical Science, Peking University Health Science Center, Beijing, China.; Human Disease Genomics Center, Peking University, Beijing, China.
* To whom correspondence should be addressed. E-mail: tpshi{at}126.com; taiping_shi@yahoo.com.cn.
In the present study, a cell-based high-throughput assay is established to identify novel human genes associated with cell viability. The assay relies on the down-regulation of Renilla luciferase (pRL) activity in a 96-well format. In addition, 2-color fluorescence probes were used to distinguish living and dead cells. As the positive control, the authors used the expression vectors encoding Bax, TNFRSF1A, and TAJ, which were widely known to effectively induce programmed cell death. They screened 409 novel genes (including alternative mRNA splicing forms) cloned in their laboratory and found that 39 genes could significantly down-regulate pRL activity. A subsequent fluorescence-based assay revealed that 4 of the 39 genes (PIP5KL1, OLFM1, RNF122, FAM26B) were associated with cell viability. Further function assays validated that the 4 genes were able to induce both necrosis and apoptosis. These results therefore indicate that a rapid and effective screening system has been developed, which should shed light on some functions of novel genes. Key Words: high-throughput screening, fluorescence, cell viability, novel gene, imaging
First published on April 28, 2006, doi:10.1177/1087057106286654 |
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