I want to study the effect of a particular transcription factor on activation and proliferation of B lymphocytes. My aim is to knock-down or knock out the transcript in primary B lymphocytes (hard-to transfect), stimulate with anti-IgM antibody and measure proliferation with thymidine incorporation assay. Please share your experiences and expert views if shRNA or CRISPR/Cas9 should be a better choice for this kind of experiment.
(Please note that I have access to P2 lab, so that's not a problem)
Thanks in advance
You can use shRNA of your TF and generate stable KD cells population. I also KD one of the TF using shRNA and got very satisfactory result. Just want to know, which profile you want to check____ mRNA, protein, or metabolite?
Thank you for your answer Iqbal Mahmud. Are you using primary B cells?
I plan to check mRNA
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