Analysis of the Effectiveness of CRISPR/Cas9-Editing of the GEX2 Gene by Ribonucleoprotein Complexes in Maize Protoplasts

The GEX2 protein is expressed in the maize gamete membranes and is necessary during fertilization at the stage of contact (adhesion) of gamete membranes. Knockout of GEX2 gene can presumably lead to impaired fertilization and, as a result, to the formation of matroclinic haploid maize embryos. The purpose of this study is to analyze the efficiency of CRISPR/Cas9 editing of the GEX2 gene after PEG-mediated transfection of maize protoplasts by ribonucleoprotein (RNP) complexes with different gRNAs. For the first time, constructs for CRISPR/Cas9 editing of the maize GEX2 gene have been created, the effectiveness of which has been proven on protoplasts and reaches 10.7%, depending on the selected gRNA and the ratio and number of components in the RNP complexes.