Do I need to insert a promotor into HDR template targeting the Rosa26 locus (mouse)

I'm not sure if this is even a reasonable question.... I want to KI a gene at the Rosa26 locus, do I need to include a promotor into the HDR template (such as CMV) or can the homology arms be designed in such a way as to utilise an endogenous promoter, if there is one?

Very grateful for any advice!


Login or Signup to leave a comment
Orchid Gnomeover 3 years ago

This is a very good question. Both options you mentioned are viable. In most cases, I would prefer adding a promoter because:

1. Simpler design

2. better control over expression level/pattern

3. In vitro test of the construct is feasible

4. ROSA26 promoter/enhancer got silenced in certain tissues

White Elfover 3 years ago

CMV promoter is not a good option as it could be silenced in vivo by methylation. You can add an additional promoter such as CAG, EF1a, etc. to enhance its expression if high expression level is what you need. However, a moderate endogenous Rosa26 promoter may be sufficient and/or beneficial in some cases.

Find your community. Ask questions. Science is better when we troubleshoot together.
Find your community. Ask questions. Science is better when we troubleshoot together.

Have a question?

Contact support@scifind.net or check out our support page.