Figure 1.

Inducible dCas9–VP64 (idCas9–VP64) system for the upregulation of the polyubiquitin gene UBC and the UBC promoter region. (A,B) Human embryonic kidney 293T (HEK293T) cells were transiently transfected with idCas9–VP64 and then treated with 5 μg/mL doxycycline (Dox) for 1 day. Before harvest, cells were treated with 10 μM NaAsO₂ for 5 h. dCas9 and UBC mRNA levels were determined by qRT-PCR (n = 3 each), normalized to GAPDH, and expressed as the fold change relative to the control (-Dox, -NaAsO₂). (C) Structure of the polyubiquitin gene UBC based on the NCBI gene reference and indication of the designed sgRNAs targeting the intron of UBC. All data are presented as means ± SEM from the indicated number of samples. *** p < 0.001 between two groups as indicated by horizontal bars. NS: Not significant.