mRNA transfection of ioGlutamatergic Neurons

This step-by-step protocol is recommended for lipid-based delivery of synthetic mRNA into ioSkeletal Myocytes in 96-well plates. It is also applicable to all ioSkeletal Myocytes disease models.

• The synthetic mRNA delivery protocol was adapted from the manufacturer's original protocol (Lipofectamine™ Stem Transfection Reagent, ThermoFisher Scientific).

• Transfection of synthetic mRNAs was tested 24h and 72h post thawing. The most robust outcomes were achieved when mRNA delivery was performed 24h after revival, therefore the 24h timepoint is recommended. The transfection efficiency was evaluated by imaging up to 18 days after mRNA delivery.

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