Dear all,
Currently, I am trying to transfect siRNA to mouse primary hepatocytes. I am wondering if anybody has some detailed protocol about the transfection.
I am using lipofectamine RNAiMAX as it is recommended to be highly efficient for primary cell transfection.
I tried to optimize the concentration of siRNA from 10, 20 and 30uM, but the efficacy is still low. I am wondering if we have to increase the concentration of lipofectamine according with the concentration of siRNA.
Thank you in advance for every advice.
Dear Redb, I can recommend to use the Lonza Nucleofector ready-to-use kit. The same protocol does work well for siRNA, gRNA, large and small DNA vector and also for CRISPR complexes. You can find more primary data and the protocol at:
https://www.lonza.com/products-services/bio-research/transfection/nucleofector-kits-for-primary-cells/nucleofector-kits-for-primary-hepatocytes/nucleofector-kits-for-mouse-or-rat-hepatocytes.aspx
Lipofectamine usually have been used for this purpose. Also you can use nano-carrier for delivery of siRNA.
hi, due to negative charge of SiRNA you can use chitosan nanoparticles for efficient delivery.
Please, check this reference citing use of Viromer BLUE (polymer-based reagent) for successful gene silencing after siRNA transfection in primary mouse hepatocytes.
Article Identification of Insulin-Responsive Transcription Factors R...
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