I would like to transfer the Dat-Cre transgene (B6 background) into a outbred strain (SW) of mice. How can this be done? Do I start with a single breeding pair (Dat-Cre B6 x SW) and then for every generation I backcross with a new SW, or do I have to start with several Dat-Cre x SW breedings? Any help is much appreciated, I am trying to understand what I need to do.
Dear Susana Lima ,
as you said, crossbreeding with animal contain this gene, is suitable.
If your animal (with Dat-Cre B6 gene) are homozygosity with this gene , all of your F1 have this gene. to get animals with homozygosity for this gene, you have two case 2:
If you have back-cross with animal ( Dat-Cre B6 gene), probably you have 50% homozygosity for this gene with 25% purity for you SW mices,
case 1: If you have backcroo with SW mice, probably you have 25% homozygot for this gene with 75% purity for you SW mice,
Cross different pairs reciprocally (autosomal control background) of males and females having Dat-Cre B6 gene with SW. The F.1 filial males and females are cyclically rotated for crossing in the next generation and repeated over a few generations till the desired level of Dat-Cre B6 gene is achieved. It must be ensured that inbreeding is avoided. Subsequently, pedigree history is maintained for further production.
lnathgubbi
Thank you very much for your help Loknath G.R. . I am trying to understand how many breeding pairs I need to start with, I have limited space. From what you wrote, I should have a relatively large number of cages.