4 replies to this topic

[mattyboyroy]

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

[Clare]

Hello

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

[mattyboyroy]

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?

[mattyboyroy]

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

Hello

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

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?

[Fungus_Dreams]

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.