overexpression system using retrovirus,without target gene sequence, in mammalian cell can affect unexpected phenotype?
for example, infected mammalian cell with pBabe-puro(control vector) can obtain some phenotype? such as ability of sphere-forming, stemness or capacity of tumorigenesis.
Hi Sang-Hun. The answer to your question (in my opinion) is "yes".
Retroviruses can integrate at multiple sites in the genome. The Moloney Leukaemia virus (which your retrovirus is based upon) is classed as a gamma-type retrovirus and these viruses preferably integrate into the promoter region of genes. Please see figure 3 in the following paper:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4866806/
Thus if your "empty" vector integrates into the promoter region of - for example - a proto-oncogene it could cause increased growth in the cell in the absence of an insert.
However this effect will probably not be noticeable in a standard transfection (unless the cells are passaged for a prolonged period) as the other transfected cells are unlikely to have the retrovirus integrate at the same place.
If you are looking at "clones" from cells transfected with empty vector this would be more noticeable.
Hope this helps!
Gary
Gary, Thank you for your comments. Always i wonder about effects of retrovirus system. as your mention, I think this problem should be considered when i use infection.
Thank you!
Sang-Hun
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