Journal
NATURE COMMUNICATIONS
Volume 11, Issue 1, Pages -Publisher
NATURE RESEARCH
DOI: 10.1038/s41467-020-14977-8
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Funding
- Biotechnology and Biological Sciences Research Council [BB/M024393/1]
- VENI Fellowship [13667]
- Dutch Research Council (NWO)
- European Union [721058]
- John Fell Fund [162/044]
- La Caixa Fellowship Grant for Post-Graduate Studies, la Caixa Banking Foundation
- Oxford University Press John Fell OUP Research Fund
- European Research Council [CANCER-RECURRENCE 648804]
- Doctor Josef Steiner Foundation
- Biological Sciences Research Council [BB/M024393/1]
- Swedish Medical Research Council
- Swedish Foundation for Strategic Research (SSF-IRC)
- BBSRC [BB/M024393/1, BB/R021422/1] Funding Source: UKRI
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Extracellular vesicles (EVs) form an endogenous transport system for intercellular transfer of biological cargo, including RNA, that plays a pivotal role in physiological and pathological processes. Unfortunately, whereas biological effects of EV-mediated RNA transfer are abundantly studied, regulatory pathways and mechanisms remain poorly defined due to a lack of suitable readout systems. Here, we describe a highly-sensitive CRISPR-Cas9-based reporter system that allows direct functional study of EV-mediated transfer of small non-coding RNA molecules at single-cell resolution. Using this CRISPR operated stoplight system for functional intercellular RNA exchange (CROSS-FIRE) we uncover various genes involved in EV subtype biogenesis that play a regulatory role in RNA transfer. Moreover we identify multiple genes involved in endocytosis and intracellular membrane trafficking that strongly regulate EV-mediated functional RNA delivery. Altogether, this approach allows the elucidation of regulatory mechanisms in EV-mediated RNA transfer at the level of EV biogenesis, endocytosis, intracellular trafficking, and RNA delivery. Extracellular vesicles (EV) facilitate intercellular transfer of biological material including RNA, but the regulatory mechanisms for their formation and transfer are incompletely known. Here the authors develop a CRISPR-based reporting system to detect the transfer of guide RNAs and identify genes not previously linked to EV-mediated RNA delivery.
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