ИСТИНА

Due to its simplicity, practicality and efficiency CRISPR–Cas9 israpidly developing as the emerging technology for genome editingon various eukaryotic systems including higher plants andanimals. Here we describe new approach for delivery of preassembledCas9–sgRNA ribonucleoproteins into dissected potato(Solanum tuberosum, cv Chicago) shoot apical meristems by chitosannanoparticles using vacuum infiltration and regeneration ofplants with edited alleles. As a model gene for genome editing,we have selected a gene encoding coilin, a major protein of Cajalbodies(CBs). In additional to its traditional role in assembly ofCBs, coilin also participates in biotic (virus attack) and abiotic(high salinity) plant stress responses. Using this method ofdelivery, we have demonstrated DNA-free genome editing andrecovery of potato plants with mutated (disrupted) coilin gene.The generated edited plants exhibit high level of resistance topotato virus Y and significant tolerance to osmotic and saltstress. This work opens up a new avenue for practical applicationof CRISPR-Cas9 technology to produce genome edited (transgene-free)potato plants and other crops. The present work wasperformed with the financial support from the Russian ScienceFoundation (grant No. 16-16-04019).