What is the best transfection reagent to use to transfect mRNA to cells/tissue in a low pH environment? I have tried most of the mRNA transfection reagents on the market & they do not work.

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Accepted Answer

The best reagent to transfect mRNA to cells is Lipofectamine® MessengerMAX™ because it has shown to further protect the mRNA molecules from degradation and has been shown to deliver the highest amount of mRNA into a wide variety of cell types. For tissues or small animal models it would be Invivofectamine® 2.0, which is a nanoparticle technology that can encapsulate and protect the payload for delivery. It can be used on cells in vitro as well as injected systemically via intravenous route or via direct injection (i.e. muscle, tumors, heart, brain). More information can be found at www.lifetechnologies.com/messengermax and www.lifetechnologies.com/invivofectamine In addition, mRNA can also be synthesized with chemically modified nucleotides to improve stability and further minimize degradation. A positive control mRNA (e.g. GFP mRNA) is also a very helpful method to optimize transfection techniques.

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When transfecting with mRNA, does the cell package the foreign RNA into vesicles reducing the efficiency of the mRNA, or are they left in the cytoplasm? If packaged away, what percent is packaged vs. what percent is left to be available for transcription? Is this observed with DNA plasmids?

We have seen quite a bit of variation in our transfection efficiency based on the number of passages prior to transfection. Do you have advice on the best passage number?

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