Extended Data Fig. 5: Validation of candidate circRNAs by the RfxCas13d/BSJ-gRNA system in cell proliferation.
From: Screening for functional circular RNAs using the CRISPR–Cas13 system
a, Heatmap display of the relative KD efficiency, cell proliferation and fold change of candidate circRNAs in HT29 (purple), 293FT (orange) and HeLa (green) cells with single BSJ-gRNA that targets each candidate circRNA. b,c, KD of circKLHL8 by RfxCas13d inhibited cell proliferation in 293FT (b) and HeLa cells (c), as revealed by MTT cell proliferation assay (middle panels) and cell confluency calculated by the surface area occupied by cells (right panels); KD efficiencies were showed on left panels. d, KD of circHIPK3 by RfxCas13d inhibited cell proliferation in HeLa cells, as revealed by MTT cell proliferation assay (middle) and by cell confluency calculated by the surface area occupied by cells (right); KD efficiencies were showed on left. e, KD of circKLHL8 by shRNAs inhibited cell proliferation in 293FT cells. KD efficiencies were showed on left; MTT cell proliferation assays were shown on right. f, KD of circKLHL8 and circHIPK3 by shRNAs in HeLa cells. g, KD of circKLHL8 or circHIPK3 by shRNAs inhibited cell proliferation in HeLa cells, as revealed by MTT cell proliferation assays. (a,b,c,d,e,f) All expression levels of RNAs were detected by qRT-PCR and were normalized to ACTB. Means ± s.d. were from three independent experiments. *: P < 0.05; **: P < 0.01; ns, not significant, two-tailed student’s t-test.
