mRNA transfection of ioMicroglia

This step-by-step protocol is recommended for lipid-based delivery of synthetic mRNA into ioMicroglia in 96-well plates. It is also applicable to all ioMicroglia 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 24 h and 96 h post thawing. The most robust outcomes were achieved when mRNA delivery was performed 96 h after revival, therefore the 96 h timepoint is recommended. The transfection efficiency was evaluated by imaging up to 10 days after mRNA delivery.

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