Hello. I try to transiently transfect CHO-K1 and HEK-293 for protein expression. To estimate ( microscopically ) the efficiency of transfection I use pAcGFP1-N1. I have tried to transfect with PEI and with Xfect. PEI was quite good for HEK-293, but gave a low level of transfection for CHO-K1 and Xfect I used only with CHO-K1 cell line and achieved a low transfection efficiency. Is there any efficient transfection reagent for both of theese cell lines and especially for CHO-K1? Thank you in advance.
Hey Voktoria,
We recently established a method for transfection with the JetMessenger transfection reagent and in vitro transcribed and immunologically silenced mRNA. This method was used for in vitro transfection of ex vivo (peritoneal macrophages), in vitro (bone marrow-derived macrophages) and MEF cells so far. But you can give it a try, if you want. We reached transfection efficiancies of 80 - 90 % with MEFs and BMDM and 65-70% with PMs.
For a detailed protocol (including primer desing, kits of choice and purification of mRNA) please have a look at:
Highly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA. J. Vis. Exp. (Pending Publication), e60143, In-press (2019).
All the best and stay healthy,
Marc
We used pCMV6-XL5 vector was purchased from the ORIGENETM for HEK 293
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