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Global analysis of human-to-mouse contact-dependent intercellular mRNA and lncRNA transfer in cell culture

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Title:Global analysis of human-to-mouse contact-dependent intercellular mRNA and lncRNA transfer in cell culture
Creators Name:Dasgupta, Sandipan and Dyagi, Daniella Y. and Haimovich, Gal and Wyler, E. and Olender, Tsviya and Singer, Robert H. and Landthaler, M. and Gerst, Jeffrey E
Abstract:Full-length mRNAs can transfer between adjacent mammalian cells via direct cell-to-cell connections called tunneling nanotubes (TNTs). However, the extent of mRNA transfer at the transcriptome-wide level (the transferome) is unknown. Here, we analyzed whole transcriptome mRNA and lncRNA transfer between heterogeneous human-mouse cell populations in in vitro co-culture using RNA-sequencing. Our data indicate that mRNA transfer is non-selective, prevalent across the human transcriptome, and that the amount of transfer to mouse embryonic fibroblasts (MEFs) strongly correlates with the endogenous level of gene expression in donor human breast cancer cells (MCF7). These results were validated by both quantitative RT-PCR and in situ hybridization, and analysis shows that typically <1% of endogenous mRNAs and lncRNAs undergo transfer. Non-selective expression-dependent RNA transfer was further validated using synthetic RNA reporters. Notably, significant differential changes in the native MEF transcriptome were observed in response to co-culture, including the upregulation of multiple cancer and cancer-associated fibroblast-related genes and pathways. Together, these results lead us to suggest that TNT-mediated RNA transfer could be a phenomenon of physiological importance under both normal and pathogenic conditions.
Keywords:smFISH, RNA-Seq, RNA Transfer, RNA Trafficking, Tunneling Nanotubes, Mouse Embryo Fibroblasts, MCF7 Breast Cancer Cells, Mammalian Cells
Publisher:Cold Spring Harbor Laboratory Press
Article Number:2021.11.28.470233v2
Date:7 March 2022
Official Publication:https://doi.org/10.1101/2021.11.28.470233
Related to:
https://edoc.mdc-berlin.de/23407/Final version

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