Delivery of CRISPR/Cas9 Ribonucleoprotein Complex into Plant Apical Meristem Cells Leads to Large Deletions in an Editing Gene

Genome editing using the CRISPR/Cas9 system is an innovation platform exploiting site-specific nuclease to create modifications (deletions or insertions) into a target region of the genome of eukaryotic organisms. Plasmid-free methods of editing via delivery of the preformed RNP complex consisting of Cas 9 endonuclease and short guide RNA are of particular interest in modern plant biology. The use of these methods rules out an integration of foreign DNA into the plant genome. We have recently developed the method of genome editing for the potato crop using delivery of the RNP editing complex immobilized on gold or chitosan microparticles into meristem cells. Here, using the gene of coilin, the structural protein of Cajal bodies which also controls plant stress resistance, we show that this method leads to the formation of large deletions of about 500 bp in the gene, which are not common for the CRISPR/Cas9 system possibly due to molecular-genetic peculiarities of meristem cells. Data presented are discussed in the context of practical applications of this method.