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-   -   conditional gene knockout (http://www.molecularstation.com/forum/knockout-mouse-forum/74938-conditional-gene-knockout.html)

ccfwang 01-13-2011 09:33 PM

conditional gene knockout
 
Hi, Everyone,

I am going to establish a conditional gene knockout mouse model. The ES cell is available in KOMP. The allele structure is showed below.
FRT--lacZ--loxP--neo--FRT--loxP--gene exon--loxP
KOMP suggest crossing with Flp mouse to remove lacZ--loxP--neo sequence first , and then crossing with Cre mouse to knockout the gene exon. However, I want to try crossing with Cre mouse to remove the gene exon.
My questions are:
1. Is it possible?
2. What problems should I take a care of?

Thanks
Lianfu Wang

Biltquirl 04-14-2011 03:17 AM

christian louboutin pumps
 
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blondeventer 05-13-2011 04:02 PM

Re: conditional gene knockout
 
Hello everyone,

I'm looking into getting a Cre line that is specific to dorsal cortex progenitors. I'm intereted in the Emx1-Cre line. Has anyone used it background other that C57/B6? Our floxed line is FVBN/Sv129. Does anyone know of a different dorsal cortex specific CRE line that works well on these background strains?

Thanks

mmorgan 06-29-2011 05:23 PM

Re: conditional gene knockout
 
Quote:

Originally Posted by ccfwang (Post 425019)
Hi, Everyone,

I am going to establish a conditional gene knockout mouse model. The ES cell is available in KOMP. The allele structure is showed below.
FRT--lacZ--loxP--neo--FRT--loxP--gene exon--loxP
KOMP suggest crossing with Flp mouse to remove lacZ--loxP--neo sequence first , and then crossing with Cre mouse to knockout the gene exon. However, I want to try crossing with Cre mouse to remove the gene exon.
My questions are:
1. Is it possible?
2. What problems should I take a care of?

Thanks
Lianfu Wang


If you cross the KOMP allele to a Cre expresser you will likely create a null allele that is a lacZ reporter. But some of these allele are "knockout first" so that they are in theory a null/reporter in the first place. The Cre excision is just a further guarantee that you have disrupted the gene product.

If you want to make a conditional allele, you need to either cross with a Flp mouse line. Or you can excise the ES cells in vitro by electroporating in a Flp plasmid.


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