Figure 3 | Scientific Reports

Figure 3

From: An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells

Figure 3

The epiCRISPR system for efficient double-gene knockout. (a) Expression of two gRNAs on the epiCRISPR vector for double-gene knockout. (b) RFLP analysis of the indel rates generated by the epiCRISPR system with gRNA multiplexed targeting DYRK1A & EMX1, AAVS1 & VEGFA and APC1 & MLH1 in hPSCs (n = 3, error bars show mean ± S.D.). (c) RFLP analysis of single cell-derived clones multiplexed targeting DYRK1A and EMX1 loci. Ctr1 is the PCR band from unmodified cells without digestion. Ctr2 is the PCR band from unmodified cells with digestion (BstXI for DYRK1A and AgeI for EMX1). Red triangles indicate the epiCRISPR-modified PCR bands; black triangles indicate unmodified PCR bands; the red asterisks indicate the homozygous knockout for both genes.

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