I have been considering to use either lentiviral or piggybac with Tet-On 3G promoter to overexpress MyoD1 in iPSC lines. I am pretty new in this stem cell research and I came to know from my former colleague that these iPSC lines are difficult to transfect and through electroporation she managed to transfect 30% with simple GFP marker of cells but it was nearly 10-15% when she tried with CRISPR vector. Does anyone enlighten me with my query that which system would be better to go for. Piggybac is simple but it depends on transfection whereas lenti should be more effective with integrating since cells will not need transfection but infection. But virus infection is how much effective for iPSCs??
In our experience (the whole institute, I mean) it is unofficially accepted that piggybac generally works better. But it may depend on your cell line or conditions. By the way, the transfection method better be electroporation or Lipofectamine.
Dear Yulia Panina,
Thank you for your reply and I am sorry to write you back late. Could you please elaborate a bit in what perspectives piggybac was better than lenti? I cam to know through One of my colleagues that someone used sleeping beauty method which is similar to piggybac and was not happy with it. Thanks in advance.
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