Hello everyone,
I am starting a project in which I'm planning to create a cell line with a specific deletion. I am going to use SUP-B15 and I need to know if anyone has ever transfected this cell line before. Do you have any suggestion how to transfect recombinant cas9 (or even plasmide encoding for cas9) and what method to use.
I have access to Neon electroporator, Lipofectamin and RNAiMax. Which one do you suggest ?
Thank you very much in advance
Hello Akram,
if you want to deliver the purified Cas9 together with gRNA as ribonucleoproteins (RNP), you can go with electroporation. Probably the best thing for these tricky cells. Alternatively, if it's too damaging for the cells and/or if you prefer using a chemical reagent, you would need the Lipofectamine CRISPR-Max, which is (I think) the same reagent as RNAiMax but with a booster (Cas9 Plus Reagent).
Another option: trying another new reagent very performant for RNP delivery, our Viromer CRISPR.
https://viromer-transfection.com/crispr
I can send you a free sample if you are motivated. As we are just launching this product, you are very welcome to give it a try with SUP-B15. Probably, the protocol needs to be adjusted for suspension cells, I can help you with this.
You can write me at this email [email protected]
Best,
Sandra
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