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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, Heyang (author)
Ghent University, Belgium,Universiteit Gent,Ghent university
Bussmann, Jeroen (author)
Leiden University, Netherlands,Universiteit Leiden (UL),Leiden University (UL)
Huhnke, Florian (author)
Max Planck Institute for Medical Research, Germany,Max Planck Gesellschaft zur Förderung der Wissenschaften e.V. (MPG),Max Planck Society for the Advancement of Science (MPG)
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Devoldere, Joke (author)
Ghent University, Belgium,Universiteit Gent,Ghent university
Minnaert, An-Katrien (author)
Ghent University, Belgium,Universiteit Gent,Ghent university
Jiskoot, Wim (author)
Leiden University, Netherlands,Universiteit Leiden (UL),Leiden University (UL)
Serwane, Friedhelm (author)
Max Planck Institute for Medical Research, Germany; Ludwig-Maximilian-University Munich, Germany; Munich Cluster for Systems Neurology, Germany,Ludwig-Maximilians-Universität München,Ludwig Maximilian University of Munich,Max Planck Gesellschaft zur Förderung der Wissenschaften e.V. (MPG),Max Planck Society for the Advancement of Science (MPG)
Spatz, Joachim (author)
Max Planck Institute for Medical Research, Germany; University of Heidelberg, Germany,Ruprecht-Karls-Universität Heidelberg,Heidelberg University,Max Planck Gesellschaft zur Förderung der Wissenschaften e.V. (MPG),Max Planck Society for the Advancement of Science (MPG)
Röding, Magnus (author)
Gothenburg University,Göteborgs universitet,RISE,Jordbruk och livsmedel,Chalmers University of Technology, Sweden; University of Gothenburg, Sweden,RISE Research Institutes of Sweden,Chalmers tekniska högskola,Institutionen för matematiska vetenskaper,Department of Mathematical Sciences
De Smedt, Stefaan (author)
Ghent University, Belgium,Universiteit Gent,Ghent university
Braeckmans, Kevin (author)
Ghent University, Belgium,Universiteit Gent,Ghent university
Remaut, Katrien (author)
Ghent University, Belgium,Universiteit Gent,Ghent university
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 (creator_code:org_t)
2021-12-16
2022
English.
In: Advanced Science. - : John Wiley and Sons Inc. - 2198-3844. ; 9:4
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  • Journal article (peer-reviewed)
Abstract Subject headings
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  • 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. © 2021 The Authors. 

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)
NATURVETENSKAP  -- Biologi -- Cellbiologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Cell Biology (hsv//eng)
NATURVETENSKAP  -- Biologi -- Utvecklingsbiologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Developmental Biology (hsv//eng)
NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Keyword

cellular uptake
mingle/single-mRNA lipoplex
protein expression
single cell
theoretical modeling
Cells
Cytology
Encoding (symbols)
Proteins
RNA
Signal encoding
Individual cells
Lipoplexes
Messenger RNA
Mingle/single-messenger RNA lipoplex
Protein expressions
Ratiometric
Single cells
Single-cell level
Liposomes
cellular uptake
mingle
single-mRNA lipoplex
protein expression
single cell
theoretical modeling
mediated delivery
endosomal escape
sirna
integrity
polyplex
vaccine
Chemistry
Science & Technology - Other Topics
Materials Science

Publication and Content Type

ref (subject category)
art (subject category)

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