RNA deep sequencing as a tool for selection of cell lines for systematic subcellular localization of all human proteins

RNA深度测序作为选择细胞系的工具,用于所有人类蛋白质的系统亚细胞定位

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作者:Frida Danielsson, Mikaela Wiking, Diana Mahdessian, Marie Skogs, Hammou Ait Blal, Martin Hjelmare, Charlotte Stadler, Mathias Uhlén, Emma Lundberg

Abstract

One of the major challenges of a chromosome-centric proteome project is to explore in a systematic manner the potential proteins identified from the chromosomal genome sequence, but not yet characterized on a protein level. Here, we describe the use of RNA deep sequencing to screen human cell lines for RNA profiles and to use this information to select cell lines suitable for characterization of the corresponding gene product. In this manner, the subcellular localization of proteins can be analyzed systematically using antibody-based confocal microscopy. We demonstrate the usefulness of selecting cell lines with high expression levels of RNA transcripts to increase the likelihood of high quality immunofluorescence staining and subsequent successful subcellular localization of the corresponding protein. The results show a path to combine transcriptomics with affinity proteomics to characterize the proteins in a gene- or chromosome-centric manner.

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