ZFIN Image: Wierson et al., 2019, FIG. 4.

Image

Figure Caption

FIG. 4.

ErCas12a activity in human HEK293T cells. (A) Schematic showing the workflow for ErCas12a and pre-crRNA expression in human cells from a cis expression plasmid. After transfection, cells are recovered and then analyzed. (B) Schematic of two pre-crRNAs designed to target AAVS1. (C) T7E1 assay showing AAVS1-pre-crRNA1 is active. Arrowheads are cleaved bands, indicating nuclease activity. (D) Schematic of four pre-crRNAs designed to target CCR5. (E) Schematic of four pre-crRNAs designed to target TRAC. (F) Graph showing percentage of indels after using pre-crRNAs to target AAVS1, CCR5, and TRAC in HEK293T cells as determined by ICE analysis. Data plot represents the mean ± SD. (G) Results displaying the wild-type and top five mutated alleles after AmpliconEZ analysis of AAVS1 targeted DNA.

Acknowledgments

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