Hello everybody,
I want to use a Cre-Lox mouse model to study the behaviour of glial cells during a Parkinson-like state. In our in vitro model, we take glial cells from newborn mice to perform our primary cultures.
I am wondering whether I can treat first these Cre-lox mouse and afterwards put newborn glial cells in culture ? Or is it better to take glial cells from Cre-Lox newborns before treating them "in vitro" in the plate with Tamoxifen ?
Thanks in advance,
Have a nice day,
Best,
Tony Heurtaux
Hi Tony,
I would treat the cells with Tamoxifen in vitro to avoid to potentially interfere with the developmental stages of the newborn mice by deleting a specific gene. FYI, we previously performed Tamoxifen treatment of primary microglial cells using a concentration of 1uM for 48h. We obtained a decrease at gene expression level of approx. 80% as measured by q-PCR.
Best,
Alessandro
Quite funny to read you on ResearchGate !
Thanks for your answer Alessandro ! See you soon ! ;-)
As far as i am concern the breeding of two parent one carrying gene to be knock out flanked by loxp and other parent carrying cre gene under the control of tissue specific promoter. The offspring therefore carry both the gene. So the gene will be knock out only the tissue that is purely determine by cre gene promoter. if your cre gene is exclusively express in in glial cell it will knock out loxP gene gene in glial cells. Therefore treatment condition is purely base on what factor determined the activate cre protein through cre gene promoter.
- Badges
- Science method