Fig. 4

Transcriptional activation with the AsCas12a variants. a Schematic of the gene activation system based on catalytic-dead AsCas12a variants. VPR, synthetic VP64-p65-Rta activation domain. b qPCR analyses of the transcriptional activation levels with the AsCas12a variants guided by a single sgRNA targeting each promoter region of IL1RN, MYOD, and HBG in human HEK293T cells and Fgf21 in mouse B16 cells, respectively. Mean values are presented with SEM, n=3 independent experiments. **p<0.01. ns, no significance. Student’s t-test, dWT-VPR sample versus corresponding dAsCas12a-VPR variant samples. c RNA-seq assessing the specificity of the dWT-, dRKA-, and dHF-VPR systems to activate IL1RN gene. d Schematic of the orthogonal gene editing and activation system based on catalytic active AsCas12a variants. e Relative mRNA expression (Activation) and genome editing efficiency (Editing) of the AsCas12a-VPR systems at MYOD and IL1RN sites with indicated lengthed sgRNAs. 15-bp sgRNAs for gene activation and 23-bp sgRNAs for genome editing. Mean values are presented with SEM, n=3 independent experiments. Indel was revealed by Deep-seq