Hi to everyone!
Does any of you have experience in transfecting siRNA in Jurkat cells and can provide a reliable method? I have both Neon transfection system and RNAiMAX Lipo, but to date none of these methods has given me any appreciable result.
Hope you can help!
Giovanni
Transfecting siRNA into Jurkat cells can be challenging. Here’s a method that might help improve your results using the Neon Transfection System and Lipofectamine RNAiMAX.
Method Using Neon Transfection System
Method Using Lipofectamine RNAiMAX
Tips:
- Optimize siRNA concentration and electroporation parameters.
- Ensure cells are healthy and in mid-log phase.
- Validate transfection efficiency with a control siRNA.
Both methods may require optimization based on your specific experimental conditions.
We have a terrible time getting anytime of nucleic acid into Jurkat cells. For a long time we tried Lipofectamine 3000 and this partially worked but never yielded an adequate efficiency. Now we only use lentivirus to introduce DNA or RNA into these cells. One things that partially helped if you have to use transfection: We applied the transfection mix and gently spin these reagents and cells in a 6-well plate (~200rcf or less) for 10min and do this repeatedly with breaks ack into the CO2 incubator to keep the cells alive. Goodluck!
Idowu Goodnews Oluwaseyi thanks for your kind answer. I'm trying NEON at the moment, using manifacturer's suggested parameters (1325V, 10ms, 3pulses). Nothing happens.
We are also considering using Lipofectamine RNAiMAX but it seems (at least from all the discussions I've red) that it yelds an even lower transfection efficiency.
Choosing T-cell leukemia cell lines for our project has not been a wise choise probably :D
Giovanni Pecoraro Did you find the solution? If not, how about this "Article Lipid Nanoparticles (LNPs) outperform Electroporation in mRN...
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