Feb 08, 2024
ASO transfection of iPSC-derived cells
- 1University College London, University of London
Protocol Citation: james.evans 2024. ASO transfection of iPSC-derived cells. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbxbonlpk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 08, 2024
Last Modified: February 08, 2024
Protocol Integer ID: 94881
Keywords: transfection, ASO, midbrain dopaminergic neurons with antisense oligonucleotide, antisense oligonucleotide, dopaminergic neuron, transfecting ipsc, aso transfection, neuron, derived midbrain, derived cell, transfection, aso
Abstract
A protocol for transfecting iPSC-derived midbrain dopaminergic neurons with antisense oligonucleotides (ASOs).
ASO transfection of iPSC-derived mDA neurons
Make up transfection mix in N2B27. The transfection mix should be 1/5 of the final volume in the well.
Transfection mix should have:
2. ASO (adjust concentration depending on cell type, ASO chemical modification, and knockdown required). Calculate final concentration required in the well not in the transfection mix.
Example - 1000 ul final volume in the well with 300 nm of ASO = 200 ul of N2B27 + 0.96 ul of DharmaFect + 3 ul of 100 uM ASO Stock
Vortex transfection mix and leave at room temperature for 30 minutes.
Aspirate media from cells and replace with the transfection mix. After adding the transfection mix add the rest of the media to the cells to make up the final volume.