Figure 1.

The single guide RNA (sgRNA) mediated CRISPR/dCas9 synergistic activation mediator (SAM) system for target gene activation. A, Schematic diagram of target gene activation, indicating that the SAM complex, consisting of three components, binds to the endogenous target gene promoter and transcriptional activation occurs. B, Three sgRNAs targeted each of the endogenous human Nr5a1, Gata4 and Dmrt1 gene promoters. TSS, transcription start site. C, Schematic representation of lenti‐dCas9‐VP64 plasmids expressing dCas9‐effector fusion proteins. D, Schematic representation of lenti‐MS2‐P65‐HSF1 (MPH) plasmids expressing the transcriptional activation complex. E, The lenti‐3βHSD‐EGFP plasmid reporter system in which 3βHSD gene promoter fragments were cloned upstream of EGFP. F, Schematic representation of the Lenti sgRNA(MS2)‐Puro vector. For the cloning of these designed sgRNAs, the insertion site of the guidance sequence lies downstream of the U6 promoter