Fig. 4.

Disruption of MYC superenhancers leads to KSHV reactivation. (A) dCas9-KRAB-sgRNA-mediated CRISPRi targeting each MYC eRNA, and dCas9-KRAB alone used as control. (B) qRT-PCR of host MYC mRNA upon CRISPR inhibition of the three MYC eRNAs. (C) shRNA-mediated RNA interference (RNAi) targeting each MYC eRNA, and scrambled sequence used as control. (D and E) qRT-PCR of MYC and viral gene mRNAs (LANA, RTA, and K2) upon shRNA transduction. (F) Immunoblot for MYC and LANA upon shRNA transduction validates the qRT-PCR results. (G) BRD4 ChIP-seq signals from the host genome during latency are aligned next to H3K27ac signals. BRD4 preferentially binds to superenhancers but not typical enhancers. (H) qRT-PCR results for MYC eRNA upon treatment with 0.5 μM of JQ1 for 24 h. (I) qRT-PCR results for MYC mRNA and viral mRNAs (LANA, RTA, and K2).