Existing methods for gene disruption are limited by their efficiency, time required for completion and potential for confounding off-target effects.
Zeb1 knockdown results in no statistically significant change in cell morphology and attempts at siRNA targeting of Abl2 and Dlc1 produced inconclusive data on cellular morphology due to confounding off-target effects (data not shown).
given that there are no confounding off-target effects – since even the most selective drugs are not absolute in their specificity.
At higher concentrations of Bay 11-7082, UPR-induced NF-κB activation was more greatly inhibited but cell death and confounding off-target effects (e.g., UPR inhibition) were also increased dramatically.
This analysis allows additional information to be gathered from the results of RNAi screening and turns potentially confounding off-target effects into a new source of information for the process being studied.
Because these results are based on eight different MYBL2-specific shRNAs, each with different knockdown efficiency, this analysis ensures the robustness of the data and eliminates potentially confounding off-target effects by individual shRNAs.
Although sometimes limited by the short efficacy period and confounding off-target effects (reviewed in Bedell et al., 2011), the use of morpholinos in zebrafish research, beginning at the turn of this century, tremendously accelerated zebrafish loss-of-function studies and solidified the relevance of investigating zebrafish orthologs of human disease genes.
The use of RNA duplexes is potentially confounded by off-target inflammatory cellular responses [ 16].
7) The conclusion that FOXO6 mediates the effects of erlotinib on SOX2 expression is not well supported by the data, because it appears that the RNAi data presented is confounded by off-target effects.
Any one-by-one comparison of a wild-type clone and a targeted clone could be confounded by an off-target effect in one of the clones.
To avoid confounding off-pathways effects of small molecules, we generated stable cells conditionally expressing a construct comprising constitutively active mDia2 fused to the membrane-associating portion of PPP1R15B (residues 1-146).
