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. 2024 Apr 8;52(8):4483–4501. doi: 10.1093/nar/gkae240

Figure 3.

Figure 3.

CCNK and CDK12 in 3′ end processing. (A) CRISPR/Cas9-mediated depletion of CCNK or CDK12 increases the mCherry/GFP ratio in HEK293 cells expressing the dual fluorescence readthrough reporter containing the distal PAS of the CPEB2 gene. 10000 cells were analyzed by FACS. (B) Depletion of CCNK or CDK12 specifically increases the expression of mCherry RNA relative to that of GFP RNA in HEK293 cells expressing the dual fluorescence readthrough reporter containing the distal PAS of the CPEB2 gene. RNA expression levels were measured by RT-qPCR using primers targeting the indicated reporter regions in cells infected with lentiviruses transducing the indicated gRNAs. (C) Depletion of CCNK or CDK12 increases RNA expression downstream of the PAS in endogenous genes. RNA expression from the MYB and YKT6 genes was measured by RT-qPCR using primers targeting the indicated gene regions in cells infected with lentiviruses transducing the indicated gRNAs. Up: upstream of PAS. Down: downstream of PAS. The above results represent two biological replicates. *P< 0.05, **P< 0.01 as compared to the gAAVS1 control.