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. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Nat Biotechnol. 2021 Jan 28;39(6):697–704. doi: 10.1038/s41587-020-00806-2

Extended Data Fig. 3. Validation of CRISPR hits via manual assays.

Extended Data Fig. 3

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.