FIGURE 1.

Expression of Cas endonucleases in Phytophthora infestans transformants. (a) Immunoblot of five representative strains transformed with the plasmid encoding the green fluorescent protein (GFP)‐tagged catalytically inactive form of the nuclease, PsNLS‐dCas9‐GFP, probed with anti‐GFP. NC1 and NC2 are negative controls, namely a strain expressing another protein and untransformed 1306, respectively. The expected size of the protein is 194 kDa. (b) Fluorescent micrograph of transformant expressing PsNLS‐dCas9‐GFP, showing localization of the protein to nuclei within a hypha. GFP, bright field, and merged channels are shown, top to bottom, with the scale bar equalling 10 µm. (c) Immunoblot of strains transformed with the plasmid encoding PsNLS‐Cas12a‐GFP, probed with anti‐Cas12a. NC is the untransformed progenitor strain. An empty lane between T9 and NC was deleted from the image. The expected size of the protein is 153 kDa. (d) Confocal image of transformant expressing PsNLS‐Cas12a‐GFP, showing a hypha on the left and a sporangium on the right. GFP, bright field, and merged channels are shown, top to bottom, with the scale bar equalling 10 µm