Hi,

I am not sure fully but just wanted to let you know.

I have observed the cytotoxicity in HL60 cells with shRNA which i tried to infect through lentivirus for genetic screen (Confirmed several times).

These screen have U6 promoter governed shRNA. What i observed that when i used this shRNA pool to infect HL-60 cells, somehow they died by 4-7 days post transfection. However 100% of infection efficiency was confirmed through RFP tag.

What we chose to do later was to use 30% infection efficiency and sort the cells through flow-cytometer.

Although your plasmid has CMV which i am not sure about, but cells are sensitive to shRNA infection.

Thank you.

Dear Santosh,

Thank you for your reply.

I have read some papers in which it is indicated that the CMV promoter (usual for many vectors, including the ones for shRNA) does not work properly in haematological-derived cells, for instance the lymphoblastoid cells.

As I have to use these cells or also the HL-60 cell line (again, haematological related) I am searching for experiences of all of you for the vector used for shRNA and the related promoters that you used.

Do you know anithing about this issue?

Roberto