I am trying to subcloning the short EF1a promoter to drive Cas9 in neurons but not sure if it works as well as CAG promoter. Anybody has experience with EF1a in neurons?
Dear Yu, I have worked extensively with a short version of the Ef1a promoter (250bp) inserted into AAV backbones, and it works great in primary neuronal cultures derived from both postnatal and embryonic rats/mice. Even in vivo it has a strong expression profile. I don’t know if it spoils anything for your applications, but besides neurons, it also expresses in glia cells. The promoter is derived from Addgene Plasmid 31481 (Certo et al, 2011 Nat Methods. Tracking genome engineering outcome at individual DNA breakpoints).
Good luck with your experiments.
Dear Yu! I have little information for undergraduates on the subject. This information may help you find the answer.
Vladimir
Dear Yu, I tried several plasmids carrying the EF-1a promoter to drive expression in embryonic primary neurons and it never work for me. Same plasmids worked perfectly in cell lines. On the other hand, once the EF-1a was replaced in the same backbone by CAG, CMV or Tubalpha-1 promoters expression in primary neurons was strongly reestablished. So I would suggest that the EF-1 promotor is not good for ectopic expression in neurons.
Hope it helps!
Dear Yu, I have worked extensively with a short version of the Ef1a promoter (250bp) inserted into AAV backbones, and it works great in primary neuronal cultures derived from both postnatal and embryonic rats/mice. Even in vivo it has a strong expression profile. I don’t know if it spoils anything for your applications, but besides neurons, it also expresses in glia cells. The promoter is derived from Addgene Plasmid 31481 (Certo et al, 2011 Nat Methods. Tracking genome engineering outcome at individual DNA breakpoints).
Good luck with your experiments.
- Badges
- Science topic
- Similar topics
- Biological Science
- Neuroscience