INCREASING EFFECTIVENESS OF CAS9 EXPRESSION USING VIRAL VECTORS

CRISPR-Cas9-based genome editing is a more accurate and easier-to-use tool than others for genome modification of plant species. Genome editing system can be benchmarked using restriction fragment length analysis by utilizing gRNA containing restriction sites and evaluating indel percentages. Transformation of binary vectors containing Cas9 and gRNA expression cassettes into Agrobacterium strains is most widely used for a wide range of plant species. Agrobacterium-mediated gene transformation is more efficient than previously shown; however, due to the inherent mechanisms of interference of foreign RNA with plant antiviral activity, Cas9 expression levels will be suppressed by the plant, reducing the efficiency of the system. A vector system based on virusescontaining the suppressor protein p19, which interferes with the plant immune system, introduced with the Cas9 protein, can improve the efficiency of genome editing based on other studies. This studyobtained a tomato bushy stunt virus (TBSV) strain TBSV_GFPΔCP with a GFP reporter gene, which can increase the expression level of the foreign gene and does not affect the symptomatology of