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. 2020 Mar 10;117(12):6571–6579. doi: 10.1073/pnas.1921786117

Fig. 1.

Fig. 1.

Improved CRISPRi for MYC-regulated noncoding genes. (A) Pooled MYCncLibrary screen using the ZNF10 KRAB domain fused to dCas9 for CRISPRi. CPM, counts per million of CRISPRi sample; CPM0, counts per million of control sample (RAMOS cells, no dCas9). (B) Schematic depiction of KRAB-mediated recruitment of the corepressor TRIM28 and histone-modifying enzymes (HMEs) to induce transcriptional repression (Upper), and co-IP experiments to confirm TRIM28 binding to dCas9-KRAB and dCas9-kPOGO (Lower). Two lanes of the blot were removed (position indicated by dotted line); the original blot is shown in the SI Appendix, Fig. S2D. IB, immunoblot; IP, immunoprecipitation; kPOGO, KRAB domain of POGK. (C) Pooled MYCncLibrary screen using kPOGO fused to dCas9 for CRISPRi. (D) Schematic depiction of SID-mediated recruitment of the corepressor SIN3A and HMEs to induce transcriptional repression (Upper), and co-IP experiments to confirm SIN3A binding to dCas9-SID (Lower). SID, SIN3A-interacting domain. (E) Pooled MYCncLibrary screen using dCas9-SID for CRISPRi in RAMOS and P493-6 cells.