A multistep high-content screening approach to identify novel functionally relevant target genes in pancreatic cancer

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Abstract

In order to foster the systematic identification of novel genes with important functional roles in pancreatic cancer, we have devised a multi-stage screening strategy to provide a rational basis for the selection of highly relevant novel candidate genes based on the results of functional high-content analyses. The workflow comprised three consecutive stages: 1) serial gene expression profiling analyses of primary human pancreatic tissues as well as a number of in vivo and in vitro models of tumor-relevant characteristics in order to identify genes with conspicuous expression patterns; 2) use of 'reverse transfection array' technology for large-scale parallelized functional analyses of potential candidate genes in cell-based assays; and 3) selection of individual candidate genes for further in-depth examination of their cellular roles. A total of 14 genes, among them 8 from "druggable" gene families, were classified as high priority candidates for individual functional characterization. As an example to demonstrate the validity of the approach, comprehensive functional data on candidate gene ADRBK1/GRK2, which has previously not been implicated in pancreatic cancer, is presented.

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Buchholz, M., Honstein, T., Kirchhoff, S., Kreider, R., Schmidt, H., Sipos, B., & Gress, T. M. (2015). A multistep high-content screening approach to identify novel functionally relevant target genes in pancreatic cancer. PLoS ONE, 10(4). https://doi.org/10.1371/journal.pone.0122946

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