In this video, we demonstrate magnetically-guided transfection of plasmid DNA in primary neuronal cell culture. Magnetofection uses an external magnetic field to guide the delivery of plasmids bound to magnetic nanoparticles into cell cytoplasm.
Protocol
1. Prepare Retroviral Construct Sub-clone the T-cell receptor (TCR) alpha and beta chains, separated by a 2A linker, into an MSCV-based retroviral vector (example, pMIAII or pMIGII). NOTE: The 2A linker allows for the stoichiometric and concordant expression of the alpha and beta TCR chains from a single open reading frame. The vector includes an IRES fluorescent protein cassette (ametrine or GFP) that is used to track transduced cells and transduction efficiency….
Divulgazioni
The authors have nothing to disclose.
Materials
DMEM, high glucose + glutamine
Corning Cellgro
10-013-CV
Dulbecco's Modification of Eagle's Medium with 4.5 g/L glucose, L-glutamine & sodium pyruvate
Retroviral Mediated Gene Transduction: A Gene Transfer Technique to Deliver a Transgene in Cultured Cells Using Engineered Retroviral Vectors. J. Vis. Exp. (Pending Publication), e20998, doi: (2023).