I've generated a conditional KO mice which could express CRE recombinase in the forebrain after postnatal day 3. Can anyone tell me that whether the primary neuron could still express Cre if I use the E16.5 embryo to isolate the primary neuron?
Yes, absolutely. If with your culture conditions you do get to a point where your cultured neurons reach a "maturation state" that is comparable to that of postnatal day 3, then they will express the Cre. It all depends on whether your culture conditions allow your neurons to mature as they do in vivo.
Agree with the previous answer. I guess your conditional CRE expression is reached through driving its expression by a specific gene promoter. In this case, it might be expressed at P3 in mice, but culture conditions can change cell commitment and differentiation quite a lot, so it's perfectly normal (at least not surprising). It might be worth testing expression of the gene driving CRE expression in your culture system to see if it's induced by your culture condition. Another thing is that conditional CRE expression is not always perfect and could be "leaky", ie CRE can be expressed outside the designed cell type / time / space.
Yes. everything is possible, even if it is against textbook knowledge. The promoters selected to drive Cre expression very often are only tested for tissue specifity in adult tissues/organs. If they have a "leaky" expression in only a few cells during a short period of embryonal development, this could easily be missed.
There are "leaky" expressions of some promotors which are difficult to predict. Collagen1a1 promoter, "sold" as being bone specific, has for instance a "leaky" expression in oocytes, that can cause deletion of a floxed-gene derived from the sperm within the first few cell stages after fertilisation.
Michael
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