Hi guys,
I'd like to know what your transfection reagent of choice is for co-transfection of miRNA 3'UTR target clone and miRNA precursor expression clone (both Lentiviral-based vectors) in suspension cells like B cell lines to assess binding specificity in Luciferase assays. Any thought and experience is appreciated.
Thanks
Hello, not sure if I got your question... but if you have the constructs in lentiviral vectors you should just pack the viruses (normally you do it in 293-derived cell lines that are easy to transfect with most methods and virus packaging plasmids) and then infect your cells with the two viruses at the same time... or am I missing something?
Hi Pietro:
Sorry, I forgot to mention that, yes the viral packaging was already done and particles are ready for co-transfection.
I see, then you don't have any other choice than mix the two viruses and co-infect (just pay attention at your terminology, transfection is used to indicate introduction of DNA into eukaryotic cells with chemical or physical methods, infection or transduction is used for viruses). Viruses are not like plasmids in bacteria, there is no selectivity for one or the other. There was a previous discussion on ResearchGate:
Good luck!
https://www.researchgate.net/post/Is_dual_infection_with_lentivirus_possible
Thanks Pietro for your helpful answer, sorry for the confusion!!, I certainly understand what you mean, I always make this mistake just in writing!!!,
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