Tet-On inducible system
Posted 08 March 2013 - 12:55 AM
After tranfecting my GOI into the cell line, I found that:
1. Gene expression is low (well, I didn't use the lentivirus system) -- makes no difference from other plasmid based expression.
2. Expression is higher when induce with higher dose of doxycycline. But at the same time, the toxicity effect of doxycycline causing the cell death.
(let's say I would like to see if my protein makes cells proliferate faster. How to measure the proliferation while a lot of cells are dying when I induce with dox?)
Not sure how many here using this system (or similar system). Do you guys encounter the problems above?
I was wondering if it is worth it to use this system as too much hurdles to overcome, balancing toxic effect and the expression level. Sigh
Posted 08 March 2013 - 05:17 AM
Posted 10 March 2013 - 07:23 PM
This is why I'm wondering. If the transfection didn't work, I shouldn't be seeing an increase in expression.
But my GOI is suppose to increase proliferation. Instead of cell growing, I see cells floating.