a-i) CRISPR assays using doxycycline-inducible Cas9 in HeLa cells. Targets are divided in ones that validated and ones that did not. For each experiment, the right-set panel is qPCR data of expression 96 hours after induction of Cas9 with doxycycline. a)
ZBTB11-AS1
b)
HP08474
c)
GREP1
d)
RP11–54A9.1
e)
G083755
f)
OLMALINC
g)
CTD-2270L9.4
h)
RP11–277L2.3
i)
ASNSD1 uORF. j-k) CRISPR assays using stably-expressing A375 Cas9 cells. j)
CTD-2270L9.4
k)
ASNSD1 uORF. For all data in this figure, n=6 technical replicates for each data point. Error bars represent standard deviation. Data was also acquired a 3 independent biological replicates based on doxycycline dose level (0.2 ug/mL, 1.0 ug/mL and 2.0 ug/mL doxycycline, as well as 0 ug/mL doxycycline). The data shown are the 1.0 ug/mL dosing level, with similar results observed for the 0.2 ug/mL and 2.0 ug/mL doxycycline dosing levels.