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JoVE Journal

Immunology and Infection

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Efficient Transfection of In vitro Transcribed mRNA in Cultured Cells Using Peptide-Poloxamine Nanoparticles

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10:16 min

August 17th, 2022

August 17th, 2022

2,992 Views

0:05

Introduction

1:19

In Vitro Transcription (IVT) of Chemically Modified mRNA

2:55

Generation of IVT mRNA/PP-sNps

5:22

Preparation of the Cells for Transfection and Transfection of the Cultured Cells

7:04

Analysis of Cell Transfection Efficacy Using Metridia Luciferase (MetLuc) Assay

7:53

Results: Analyzing the Transfection Efficiency of MetLuc-mRNA/PP-sNp

9:12

Conclusion

필기록

Peptide-poloxamine nanoparticles is one of the potential pulmonary mRNA delivery systems. Our protocol makes the peptide-poloxamine nanoparticles with higher productivity and efficiency, which facilitates the development of the mucosal mRNA vaccin

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A self-assembled peptide-poloxamine nanoparticle (PP-sNp) is developed using a microfluidic mixing device to encapsulate and deliver in vitro transcribed messenger RNA. The described mRNA/PP-sNp could efficiently transfect cultured cells in vitro.

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