Hello all,
I would like to knockdown some gene in mouse bone marrow derived mast cell to produce a stable expressing. Could you suggest me an appropriate method to do?
After knockdown, I also want to perform beta-hexosaminidase assay. I'm afraid that knocking down could affect the cell sensitivity...
Thank you for your help.
Hi Rania,
You can knockdown a gene with various techniques, such as using siRNA or shRNA against the gene of your interest. You could pack shRNA against the gene in question and have it packaged into lentivirus (abmgood has one of the best kits and works like a charm!) and can treat your cells with the virus. Depending on the funding available, you can buy ready to use shRNA particles (expensive) or make your own using shRNA lentiviral vectors (preferred) from open biosystems (This was part of Thermo, now taken over by GE). Not sure how much impact it has on the pricing now, as GE dharmacon products are kind of expensive!!
You will need special approval for making this lentivirus from your university agency which handles the regulatory compliance with various biohazard materials and infectious pathogen handling. I suggest you also purchase a virus titer kit, so you know how much virus you are using to knockdown the gene.
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