Fig. 3: Cas9d-MG102-2 is a highly active nuclease in human cells.
From: Compact Cas9d and HEARO enzymes for genome editing discovered from uncultivated microbes
a Flow diagram of Cas9d-MG102-2 mRNA delivery to K562 cells. The sgRNA with a 20 or 24 bp guide and the human codon-optimized nuclease mRNA were nucleofected into cells, genomic DNA was extracted and editing at the expected target site was determined via amplicon NGS. b Nuclease activity was evaluated for the Cas9d-MG102-2 nuclease at two targeting sites in the TRAC locus (guides A1 and B1) with increasing concentrations of sgRNA (150, 300, and 450 pmol/reaction). The mock control represents background editing levels at the target region in the absence of mRNA and guide.
