What is the best protocol to induce Cre recombinase via tamoxifen in bone marrow derived macrophages?

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Manoj Balakrishna Menon

We also have similar problems that the toxicity is high, but the deletion is minimal. I still dont have a solution, but the general conclusion from literature/discussions was that the efficiency differs too much depending on the locus. Anyway i could induce better deletion, when cells are in better dividing state and once confluent there was hardly any effect. Are you treating after complete differentiation??

Elvira Mass

Currently we are trying after complete differentiation. Before, we added 4-OHT at day 4 during the differentiation.

I tried 1 and 2 uM. If I use less then the recombination efficiency is very low.

Francisco Javier Rodríguez Baena

I was wondering if you use the tamoxifen ex-vivo or not. I guess you isolate the BMDM from your mice and then treat them with 4-OHT? Would be possible to treat the animals with 4-OHT and then isolate the BMDM? I haven't tried yet but I am about to put my hands on this and I was wondering if this may works. Best.