I'm interested in using baculovirus (BacMan system) mediated delivery during my project involving store operated Ca2+ entry (SOCE) and STIM/ORAI signal transduction. Baculovirus-transfection would be used to over-expression of genes of interest within HEK293, CHO and MDA MB 435s cell lines. I'm aware several labs used this system primary to purify STIM and ORAI proteins. I'm interested to use it during measurement of Ca2+ in the population of cells. The focus of my project is both pharmacology and signalling transduction of ORAI channels. My current worry is: Would I be able to draw valid conclusions about signalling transduction if I'm using baculovirus-transfection?
The only specific concern with viral transduction is whether that by itself causes an effects such as altering calcium stores. To control for this, you definitely should transduce cells with empty virus lacking a gene for overexpression (or an irrelevant gene). As long as virus with your gene of interest does something significantly different than empty virus, you should be able to draw a conclusion. Perhaps you should first look at calcium flux in cells with or without control virus just to make sure there won't be any huge issues later. Otherwise, you are basically planning to study your gene of interest by overexpressing it in transformed cell lines. An enormous amount of important information on signal transduction has been obtained this way, but people will also question the physiological relevance. Whether it's a good system really just depends on the particular question you want to address and the stage of the project. For example, you might learn a lot about a drug and then later test it in animals. Good luck.
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