Producing a double knockout mouse - double knockout mice (Feb/02/2010 )

Hi all,

I am after some help regarding the production of a double knockout mouse colony (DKO), basically we have 2 different colonies of mice, each as a different gene knocked out eg, colony 1 APOE (kO) & colony 2 CD31 (KO), (both are C57/Bl6 background) and there are no viability or fertility problems with either mouse colony.

In order to produce a Double knock out mouse colony is it best to:

1) mate a Het (-/+) APOE mouse (CD31 +/+), with a Het (-/+) CD31 mouse (APOE +/+)

2) mate a Knockout (-/-) APOE mouse (CD31 +/+), with a Het (-/+) CD31 mouse (APOE +/+) or Vice versa.

3) mate a knockout (-/-) APOE mouse (CD31 +/+) with a knockout (-/-) CD31 mouse (APOE +/+)?

I have been doing a bit of reading and have got very confused, any help and advice would be greatly appreciated.



regards

MBR

Hello

If there are no fertility/viability issues with the single KOs why don't you just breed the 2 KOs together?
Clare

I should of said i realise that 1+2 would result in the production of hets, the idea being that i could then breed the hets to produce possible double knockouts

is this a better idea than just mating the apoe knockout with the CD31 knockout?

Clare on Feb 2 2010, 12:21 PM said:

I assumed i should do this however upon reading various forums they advise that you should mate hets in single knockouts, does the same apply for double knockouts?

Well for starters, what genotypes do you need for your experiments, and how many animals of each genotype do you need?

For example, a common general technique is to use, say, 24 experimental animals (-/-) and 24 control animals (+/-). Both experimental and control mice are produced from the animals you're trying to pair now. Some people also use the full wildtype pups, +/+, depends on your experiment.