Hi everyone,
I am trying to transfect the GFP mRNA with dendrimers with 1:10 charge ratio. However I am unable to achieve the significant transfection as compared to lipofectamine. Please suggest the necessary inputs.
Thanks and regards
Dear Pooladanda,
we also tried Lipofectamine, but never could achieve transfection or translation of mRNA for GFP or any tested protein. Viromer substances or Lipofectamine either did not work or killed the cells (in our case macrophages).
Moreover, every attempt to transfect DNA led to macrophage death and nothing was expressed.
However, we recently established a method for transfection with the JetMessenger transfection reagent from Polyplus and in vitro transcribed and immunologically silenced mRNA.
For a detailed protocol (including GFP as an example) please have a look at:
Article Highly Efficient Transfection of Primary Macrophages with In...
It worked for peritoneal macrophages, BMDM and MEFS with transfection efficancies between 65% (PMs) and 90% (BMDM and MEFs).
I hope this was helpful to you.
All the best and stay healthy,
Marc
Thank you so much Mark for your prompt response. Actually, my aim of experiment to deliver the mRNA through dendrimers. Let's see others experience.
Thanks again
- Badges
- Science method