1. High efficiency of gene deletion in all tested genetic backgrounds of A. fumigatus.
High efficiency of gene deletion in all tested genetic backgrounds of A. fumigatus. In vitro-assembled Cas9 RNPs coupled with microhomology-mediated integration of the HygR cassette were tested in ΔakuB (A), Af293 (B), and DI (C) strains. (Above) Representative transformation plates are shown for each strain using 2 µg of the HygR repair template that is flanked by 35-bp microhomology arms. (Below) The assessment of pksP deletion efficiency across different strains is plotted as the number of ΔpksP mutants out of the total number of transformation colonies. Deletion efficiencies were assessed based on the color of conidia. The ΔpksP mutant produces white colonies, while ectopic integrations result in green colonies. Deletion efficiencies represent the average from at least three independent transformations. Error bars represent the standard deviation calculated for each combination of strain, the size of HygR microhomology arms, and concentration of the HygR repair template for all experimental replicates.
Qusai Al Abdallah et al. mSphere 2017; doi: /mSphere