I am trying to express a short RNA sequences in the nucleus in Drosophila. Are there any UAS-based expression vectors in flies that do not have PAS in the 3'? Or on the other hand, is it possible to just express a transcription termination sequence at the 3' of my transcript so that my transcript terminates before reaching the PAS? I don't want to have to mess with the vector backbone itself (removing PAS...etc.). I am quite new to molecular work regarding vectors, so if I am missing something important, under a misunderstanding of sorts, or if you have good suggestions, please let me know. Thank you!
The optimal way to express such constructs would be to add a self cleaving ribozyme directly downstream of where you want the transcript to terminate. A HDV ribozyme should do the trick.
Talk with folks in your lab. Just because you want to do something doesn't mean it's possible or practical. Do what you know will work.
As my Ph.D advisor said "you can't rush quality".
Hanna Alalam Thanks for the suggestion. I thought about that, but couldn't find a specific sequence that may work in Drosophila. In any case, I did find a vector (gRNA vector) that expresses via UAS but has no polyA signal in the C-term. Thanks!
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA