Hi,
What are the functional assays which I can perform to investigate the role of genes after knocking them down or potentially over expressing some genes of interest in cell lines?
Kindly help.
Hi Sumera,
In vitro functional assays such as cell proliferation/cytotoxicity, colony formation, invasion and migration, soft agar assay, wound/scratch healing, apoptotic assays, sphere formation assays etc. you can perform followed by exogenous manipulation of the gene(s). The type of assays depends on the functionality of your gene of interest.
Best of luck
Farhad
I agree. Standard assays are cell proliferation, toxicity, migration, invasion assays. That is when you have no idea about the function of your gene. Well, guilt-by-association principle might be very helpful in guessing function of your gene/protein. Check in databases which proteins interact with your protein of interest. Look at tissue-specific expression of protein. The you could use appropriate cell lines. Also check for genes , which are co-expressed with the gene of your interest, i.e. share common pattern of expression, by looking at public databases. Look also if the mutations/SNPs in your gene are associated with some disorders. That might be helpful to design appropriate functional assay for your gene of interest.
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