Help from any mouse geneticists!

[Jet915]

I need some major help on a problem. Im taking a neurobiology grad. course and need help on a question. Im more of a physiologist by the way so I know nothing about this. Here is the question.

Describe how to make an inducible, reversible and cell-type-specific knockout mouse (all in one animal). Show the arrangement of any targeting constructs , the wild-type and targeted loci, and transgenes to be used. Describe the steps from manipulating the ES cells to generation of the mice to the breeding schemes used to obtain the final animals and describe how the knockout is regulated.

What the hell?!?!? Unfortunately, the only reason I am taking this class is cause there are no physiology courses offered here this quarter. Answer can only be half a page too...............HELP!

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[AegisZero]

Are you sure he means knockout? Does RNA interference or blockage of translation count?

Knockout in a technical sense means you mess up the gene in a highly specific manner. I dont think that is what he is asking.

If he IS asking about gene silencing, or stopping the translation of a specific protein, then I think I understand a question. If this is the case IM me on AIM at AegisZero

 

[AegisZero]

1. Put a tetracyline promoter to control the Cre-activating ligand. Can control as a tet-on/tet-off system (by use of tetracycline)

2. Put a tissue specific promoter in front of Cre protein-ligand dependent (LC).

3. Insert lox sites into the gene you wish to knock out


If he means reversible in the sense that you can stop it, the above is correct. otherwise, you need to do RNAi. You would essentially have to control the proteins Dicer and whichever proteins actually mediate RNAi.