Hello. I am trying to transduce the cHL and NHL cell lines (L428, Ca46, Ramos, U2932) with lentivirus with expression vector CD550a1 with miRNA insert. The transduction efficiency is very low but I managed to select positive cells with puromycin. However I do not observe the expression of my miRNA - no difference between the expression vector and the empty vector. When I performed rt qpcr on HEK cells used to produce the virus there was also no difference in expression between expression vector and control. The cells express GFP and are resistant to puromycin, but no miRNA expression. I tried pLV and pcdh vectors, insert has around 200 bp flanks (total 500 bp). What could be the reason and what to do to fix it?
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Methods
- Cloning
More Greta Sawicz's questions See All
Similar questions and discussions