Liposome Delivery of Fluorescent-Labeled Biomolecules

FBS Superior, dexamethasone, 3-isobutyl-1-methyxanthine, insulin, and pioglitazone were provided by Astra Zeneca. P/S, transferrin Alexa Fluor 568 (T23365; Invitrogen), EGF Alexa Fluor 488 (E13345; Invitrogen), LDL-pHrodo Red (L34356; Invitrogen), LDL-Alexa Fluor 488 (homemade) were as previously reported (Franke et al., 2019 (link)). pH calibration buffers were from Invitrogen (P35379). LNPs were prepared using the cationic ionizable lipids MC3, L608, ACU5, ACU22, MOD5, and L319 (AstraZeneca) and the helper lipids cholesterol (Sigma-Aldrich), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC; CordenPharma), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N [methoxy (polyethylene glycol)-2000] (DMPE-PEG2000; NOF Corporation), and contained CleanCap eGFP mRNA (5-methoxyuridine, cat. no. L-7201) and/or CleanCap Cy5 eGFP mRNA (5-methoxyuridine, batch no. WOTL18871, cat. no. L-7701; TriLink Bio Technologies). Formaldehyde was from Merck and digitonin from Sigma-Aldrich.

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Paramasivam P., Franke C., Stöter M., Höijer A., Bartesaghi S., Sabirsh A., Lindfors L., Arteta M.Y., Dahlén A., Bak A., Andersson S., Kalaidzidis Y., Bickle M, & Zerial M. (2021). Endosomal escape of delivered mRNA from endosomal recycling tubules visualized at the nanoscale. The Journal of Cell Biology, 221(2), e202110137.

Publication 2021

insulin pioglitazone transferrin Alexa Fluor 568 EGF Alexa Fluor 488 cholesterol DMPE-PEG2000 Formaldehyde digitonin

5 methoxyuridine Alexa 568 Alexa fluor 488 Buffers Cationic Cholesterol Dexamethasone Digitonin Distearoyl sn glycero 3 phosphocholine Dmpe Formaldehyde Insulin Lipids Methoxy polyethylene glycol Mrna Phosphoethanolamine Pioglitazone Transferrin

Corresponding Organization : Max Planck Institute of Molecular Cell Biology and Genetics

Other organizations : AstraZeneca (Sweden), AstraZeneca (United States)

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Variable analysis

independent variables

FBS Superior
Dexamethasone
3-isobutyl-1-methyxanthine
Insulin
Pioglitazone
LNPs prepared using the cationic ionizable lipids MC3, L608, ACU5, ACU22, MOD5, and L319 (AstraZeneca) and the helper lipids cholesterol (Sigma-Aldrich), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC; CordenPharma), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N [methoxy (polyethylene glycol)-2000] (DMPE-PEG2000; NOF Corporation), and contained CleanCap eGFP mRNA (5-methoxyuridine, cat. no. L-7201) and/or CleanCap Cy5 eGFP mRNA (5-methoxyuridine, batch no. WOTL18871, cat. no. L-7701; TriLink Bio Technologies)

dependent variables

Measured outcomes not explicitly mentioned

control variables

Transferrin Alexa Fluor 568 (T23365; Invitrogen)
EGF Alexa Fluor 488 (E13345; Invitrogen)
LDL-pHrodo Red (L34356; Invitrogen)
LDL-Alexa Fluor 488 (homemade)
PH calibration buffers (P35379; Invitrogen)
Formaldehyde (Merck)
Digitonin (Sigma-Aldrich)

positive controls

Not specified

negative controls

Not specified

Annotations

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