In vivo multiplexed gene activation via improved Cas12a

To evaluate the specificity of CRISPR activation by vgdCas12a on a genome-wide scale, we carried out whole-transcriptome RNA-seq of HEK293T cells with the TRE3G-GFP reporter transfected with either WT dCas12a or vgdCas12a combined with the TRE3G-targeting crRNA. We also included a non-targeting crRNA as negative control for each case. Total RNA was extracted from cells transfected with Tet or LacZ crRNA, and with dCas12(WT) or vgdCas12, and library preparation and next-generation sequencing were performed by Novogene