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  Together is better: mRNA Co-encapsulation in lipoplexes is required to obtain ratiometric Co-delivery and protein expression on the single cell level

Zhang, H., Bussmann, J., Huhnke, F. H., Devoldere, J., Minnaert, A.-K., Jiskoot, W., et al. (2021). Together is better: mRNA Co-encapsulation in lipoplexes is required to obtain ratiometric Co-delivery and protein expression on the single cell level. Advanced Science, 2102072, pp. 1-14. doi:10.1002/advs.202102072.

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 Urheber:
Zhang, Heyang, Autor
Bussmann, Jeroen, Autor
Huhnke, Florian H.1, Autor           
Devoldere, Joke, Autor
Minnaert, An-Katrien, Autor
Jiskoot, Wim, Autor
Serwane, Friedhelm1, Autor           
Spatz, Joachim P.1, Autor           
Röding, Magnus, Autor
Smedt, Stefaan C. De, Autor
Braeckmans, Kevin, Autor
Remaut, Katrien, Autor
Affiliations:
1Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_2364731              

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Schlagwörter: cellular uptake, mingle/single-mRNA lipoplex, protein expression, single cell, theoretical modeling
 Zusammenfassung: Liposomes can efficiently deliver messenger RNA (mRNA) into cells. When mRNA cocktails encoding different proteins are needed, a considerable challenge is to efficiently deliver all mRNAs into the cytosol of each individual cell. In this work, two methods are explored to co-deliver varying ratiometric doses of mRNA encoding red (R) or green (G) fluorescent proteins and it is found that packaging mRNAs into the same lipoplexes (mingle-lipoplexes) is crucial to efficiently deliver multiple mRNA types into the cytosol of individual cells according to the pre-defined ratio. A mixture of lipoplexes containing only one mRNA type (single-lipoplexes), however, seem to follow the “first come – first serve” principle, resulting in a large variation of R/G uptake and expression levels for individual cells leading to ratiometric dosing only on the population level, but rarely on the single-cell level. These experimental observations are quantitatively explained by a theoretical framework based on the stochasticity of mRNA uptake in cells and endosomal escape of mingle- and single-lipoplexes, respectively. Furthermore, the findings are confirmed in 3D retinal organoids and zebrafish embryos, where mingle-lipoplexes outperformed single-lipoplexes to reliably bring both mRNA types into single cells. This benefits applications that require a strict control of protein expression in individual cells.

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Sprache(n): eng - English
 Datum: 2021-10-272021-05-182021-12-16
 Publikationsstatus: Online veröffentlicht
 Seiten: 14
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1002/advs.202102072
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Titel: Advanced Science
  Andere : Adv. Sci.
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: Weinheim : Wiley-VCH
Seiten: - Band / Heft: - Artikelnummer: 2102072 Start- / Endseite: 1 - 14 Identifikator: ISSN: 2198-3844
CoNE: https://pure.mpg.de/cone/journals/resource/2198-3844