Hello,
I recently came upon this phrase in the Addgene webpage describing lentiviral transduction.
"Typically cells transduced with lower dilutions of the virus will have higher levels of expression. Consider expanding populations transduced with a variety of dilutions and pick the population that has the most desirable level of expression."
I couldn't understand this, as to my thoughts, higher dilutions of the virus should result in bigger MOIs (more lentiviral particles and more lentiviral genome copies integrated per each cell) and thus should result in higher expression. I am currently troubleshooting lentiviral transduction to BJ fibroblasts, trying out different viral titers.
Could anyone give me an explanation on this?
Thank you.
It's a balance between expression level and cell viability/cell death. Too much transduction can reduce cell viability (even without death) and could result in lower expression due to cellular stress.
As Addgene described, there will be a sweet spot which balances the two aspects of transduction.
the higher moi of lv, the more lv enter into the same single cell, the transduce/integration rate might lower (interfered by too many genes)?
High quantities of virus could lead to cytotoxic side effect,therefore in some cases it may be better choose a artifacts due to non^ healthy cells.
MOI for Lenti virus range from 1 to 30.MOI describe number of virus particles needrd to infect one cell according to special equation for more detailes see attached ref.
https://ibidi.com>support.
Jordan R. Yaron Thank you so much for your reply, Jordan!
I haven't thought about the possibility of cells being less viable without obvious cell death, and it makes complete sense to me.
Indeed, when I transduce the cells with higher virus concentrations, a portion of the cells die off and the cells that have survived show very slow proliferation.
Thanks again, and I'll keep trying on finding that sweet spot between cell stress and protein expression.
Junjie Shao That could also be a possibility!
Thank you very much for your answer, Junjie.
Salameh Barhoom Thank you for your answer, Salameh!
I would definitely check out the reference at iBidi.
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