Cell-type Specific Ribosomal Tagging Allows for Simultaneous Multi-Tissue Translatomic Analysis
Eng, J.; Youn, S.-w.; Kitajewski, J.
Show abstract
In vivo transcriptomic analysis has advanced significantly with the development of single cell technologies. However, bulk RNA sequencing also continues to provide information on critical signaling pathways and cellular responses. The isolation of mRNA by polysome immunoprecipitation can identify genes undergoing active translation. Unfortunately, the inability to profile multiple cell types from the same sample remains a major downfall of the technique and limits broader analysis of the tissue microenvironment. In this study, we demonstrated the feasibility of immunoprecipitating polysome-associated mRNA from different cell types by strategically expressing differently tagged Rpl22 subunits. Using this technique, we isolated two distinct sets of high quality transcripts from intact B16F10 melanomas, endothelial cells and B16F10 tumors cells, for further molecular analysis.
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