Off-target effects recur in 3/4 different cell lines and are independent of preparation protocols. (a) After transfection of the siRNA directed against GFP, expression of the genes CYLD and SOAT was measured with qPCR in different cell lines after 48 h. As negative controls, a siRNA directed against a candidate tumour supressor gene (siRNA1) was transfected and a mock transfection was performed. Values are normalized to mock and the median of 2 housekeeping genes. (b) 2 different siRNA molecules with the same target sequence but of different methodological origin were transfected, one generated by in vitro transcription (see Methods) and a second siRNA was obtained by oligonucleotide synthesis (Table 1). After transfection, the expression of CYLD and SOAT was measured in HEK and HeLa cells and normalized expression after mock transfection and the median of 2 housekeeping genes (PGK, DCTN2).
Tschuch et al. BMC Molecular Biology 2008 9:60 doi:10.1186/1471-2199-9-60