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A CRISPR-Cas9-based...
A CRISPR-Cas9-based reporter system for single-cell detection of extracellular vesicle-mediated functional transfer of RNA
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de Jong, OG (author)
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Murphy, DE (author)
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Mager, I (author)
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Willms, E (author)
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Garcia-Guerra, A (author)
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Gitz-Francois, JJ (author)
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Lefferts, J (author)
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- Gupta, D (author)
- Karolinska Institutet
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Steenbeek, SC (author)
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van Rheenen, J (author)
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- El Andaloussi, S (author)
- Karolinska Institutet
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Schiffelers, RM (author)
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Wood, MJA (author)
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Vader, P (author)
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(creator_code:org_t)
- 2020-02-28
- 2020
- English.
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In: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 11:1, s. 1113-
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Abstract
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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.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Medicinsk genetik (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Medical Genetics (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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de Jong, OG
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Murphy, DE
-
Mager, I
-
Willms, E
-
Garcia-Guerra, A
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Gitz-Francois, J ...
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show more...
-
Lefferts, J
-
Gupta, D
-
Steenbeek, SC
-
van Rheenen, J
-
El Andaloussi, S
-
Schiffelers, RM
-
Wood, MJA
-
Vader, P
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show less...
- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Cell and Molecul ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Medical Genetics
- Articles in the publication
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Nature communica ...
- By the university
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Karolinska Institutet