Similarly, RNAi has temporary knockdown effects, unpredictable off-target influence and too much background noise [ 1].
Furthermore, the abundance of the antigen of interest relative to off targets influences the outcome.
ASA has many off-target effects that influence the inflammatory response including inhibition of NFκB [32], augmentation of SOCS-2 and decrease of TRAF6 expression [33], [34].
High concentrations of fasudil most likely result in off-target effects and influence the function of other kinases, as has been demonstrated for protein kinase N1 (PKN1) and protein kinase C (PKC) (Davies et al., 2000; Lingor et al., 2007).
It is not known whether these off-target effects of tamoxifen influence the fate of the cells in lineage-tracing experiments in the mammary gland.
The doses commonly used by researchers to obtain recombination in the mammary epithelium tend to be higher than those used for other organs, and the concern arises that these high doses of tamoxifen may have unrecognized off-target effects that may influence the fate of the cells under study.
To consider the potential influence of off-target gene modifications (Yee, 2016), we detected the most probable ten off-target modification sites which were designed at the CRISPR website (http://crispr.mit.edu/) (Supplemental Table. 3) and found that there was no off-target gene modification (Supplemental Fig. 2), which excluded the potential influence of off-target gene modifications.
Further, to consider the potential influence of off-target gene modifications (Yee, 2016), we detected the most probable ten off-target modification sites which were designed at the CRISPR website (http://crispr.mit.edu/) (Table S3) and found that there was no off-target gene modification (Fig. S2), which excluded the potential influence of off-target gene modifications.
All in all, these data indicated that the phenotypes in these Clock knockout mESCs should mainly be due to the loss of Clock but not the influence of off-target gene modifications or loss of other Clock functions.
To consider the potential influence of off-target gene modifications (Yee, 2016), we detected the most probable ten off-target modification sites which were designed at the CRISPR website (http://crispr.mit.edu/), whose off-target cleavage site sequences and gene numbers were as follows in Table S3.
