conditional gene knockout

[ccfwang]

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]

hello
i'm new to here
glad to see all

[blondeventer]

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]

Originally Posted by ccfwang

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.