In line with previous data suggesting low efficacy of the CMV promoter in some cell lines (Qin et al. PlosOne 2010), I experience low transcriptional activity in a neuroblastoma cell line after lentiviral transduction of a pCDH-CMV-MCS-EF1-Puro vector and selection. My question is whether it would be worth the effort to isolate individual cell clones with the aim to specify cells with higher CMV driven expression than the average? In other word, in general, is there a variation in CMV activity in between individual cells after lentiviral transductions?
Yes, there will be a lot of variation introduced by where the vector has integrated. For example a copy integrated near a strong cis enhancer will give that clone a bright signal. Alternatively, an integrant may experience silencing (happens quite a bit with CMV) so little or no expression. Why choose CMV? Perhaps for neuroblastoma, a strong neuronal promoter (e.g. synapsin) might do?
The CMV promoter in pCDH is a shrunken CMV,which often doesn't express in a lot of cell lines.I suggest try another vector.
Thank you for constructive feedback. I appreciate it! By analysis of clones I concluded that the pCDH vector do work, but in a cell and cDNA specific manner.
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