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Testing the In Vitro and In Vivo Efficiency of mRNA-Lipid Nanoparticles Formulated by Microfluidic Mixing
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Bioengineering
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JoVE Journal Bioengineering
Testing the In Vitro and In Vivo Efficiency of mRNA-Lipid Nanoparticles Formulated by Microfluidic Mixing

Testing the In Vitro and In Vivo Efficiency of mRNA-Lipid Nanoparticles Formulated by Microfluidic Mixing

DOI:
10.3791/64810-v

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08:55 min

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January 20, 2023

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, , , ,
1Department of Bioengineering,University of Pennsylvania

Chapters

  • 00:05Introduction
  • 00:50Pre-Formulation Preparation
  • 01:51Preparation of the Lipid and Nucleic Acid Mixes
  • 02:33Microfluidic Formulation of mRNA-LNPs
  • 03:56Post-Formulation Processing and Characterization of the mRNA-LNPs
  • 04:51In Vitro Transfection of HepG2 Cells
  • 05:55In Vivo Evaluation of mRNA-LNPs in Mice Following Tail Vein Injection
  • 07:34Results: Formulation of mRNA-LNPs and the Assessment of In Vitro and In Vivo Performance
  • 08:24Conclusion

Summary

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Here, a protocol for formulating lipid nanoparticles (LNPs) that encapsulate mRNA encoding firefly luciferase is presented. These LNPs were tested for their potency in vitro in HepG2 cells and in vivo in C57BL/6 mice.

Tags

MRNA-lipid NanoparticlesMicrofluidic MixingLipid Nanoparticle FormulationIn Vitro TestingIn Vivo TestingLipid ComponentsMRNA DilutionMicrofluidic InstrumentSyringe PreparationPBS DilutionCitric Acid BufferLipid Stock SolutionsIonizable LipidHelper LipidCholesterolLipid-anchored PEG

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