Transcriptomic profiling of HBG gene expression in HEK293T cell line
Efficiency of HBG gene activation by the Cas12f/cgRNA system. In a doxycycline-inducible dCas12f-VPR knock-in (KI) HEK293T cell line, transiently transfected plasmids encoding U6, linear, and circular gRNAs targeting HBG1, and cgRNAs targeting mNeonGreen (negative control, NC) to activate HBG gene expression. RNA-seq analysis showed that cgRNAs exhibited increased efficiency in the target gene HBG1 and slight lower specificity when compared with U6 and linear gRNAs. Overall design: To define whether Cas12f/cgRNA system could enhance dCas12f-based endogenous gene activation, we transiently transfected different plasmids encoding gRNAs to activate HBG gene expression in the doxycycline-inducible dCas12f-VPR knock-in (KI) HEK293T cell line. Then we performed RNA-sequencing and compared our date with HBG gene expression from publicly available databases.