I intend to generate immortalized BMDM knockouts through CRISPR/Cas9 using the pX330 plasmid from Zhang Lab, and my best option of transfection so far would be electroporation through the Lonza Nucleofector 4D equipment. Does anyone know the best reagent/kit to use for these particular cells in this equipment? Would the kit for primary BMDM work just as well?
If you know about any plasmid transfection methods other than electroporation that have also worked well for immortalized BMDM, please let me know. Thank you!
Elisa, I believe that this thread is a good starting point to answer both of your questions.
https://www.researchgate.net/post/siRNA_in_bone_marrow_derived_macrophages_BMMs
Lipofectamine-based transfection should work for these cells although the efficiency will be lower:
http://www.bio-protocol.org/e28
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