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A high-throughput G...
A high-throughput Galectin-9 imaging assay for quantifying nanoparticle uptake, endosomal escape and functional RNA delivery
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- Munson, Michael J. (författare)
- AstraZeneca AB
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- O’Driscoll, Gwen (författare)
- AstraZeneca AB
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- Silva, Andreia M. (författare)
- AstraZeneca AB
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- Lázaro-Ibáñez, Elisa (författare)
- AstraZeneca AB
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- Gallud, Audrey, 1988 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Wilson, John T. (författare)
- Vanderbilt University
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- Collén, Anna (författare)
- AstraZeneca AB
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- Esbjörner Winters, Elin, 1978 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Sabirsh, Alan (författare)
- AstraZeneca AB
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(creator_code:org_t)
- 2021-02-16
- 2021
- Engelska.
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 4:1
- Relaterad länk:
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https://research.cha... (primary) (free)
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https://www.nature.c...
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https://research.cha...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- RNA-based therapies have great potential to treat many undruggable human diseases. However, their efficacy, in particular for mRNA, remains hampered by poor cellular delivery and limited endosomal escape. Development and optimisation of delivery vectors, such as lipid nanoparticles (LNPs), are impeded by limited screening methods to probe the intracellular processing of LNPs in sufficient detail. We have developed a high-throughput imaging-based endosomal escape assay utilising a Galectin-9 reporter and fluorescently labelled mRNA to probe correlations between nanoparticle-mediated uptake, endosomal escape frequency, and mRNA translation. Furthermore, this assay has been integrated within a screening platform for optimisation of lipid nanoparticle formulations. We show that Galectin-9 recruitment is a robust, quantitative reporter of endosomal escape events induced by different mRNA delivery nanoparticles and small molecules. We identify nanoparticles with superior escape properties and demonstrate cell line variances in endosomal escape response, highlighting the need for fine-tuning of delivery formulations for specific applications.
Ämnesord
- NATURVETENSKAP -- Biologi -- Cellbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Cell Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Farmakologi och toxikologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Pharmacology and Toxicology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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