Leadzyme formed in vivo interferes with tobacco mosaic virus infection in Nicotiana tabacum

Wyszko, Eliza; Nowak, Monika; Pospieszny, Henryk; Szymanski, Maciej; Pas, Jakub; Barciszewska, Mirosława Z.; Barciszewski, Jan

We developed a new method for inhibiting tobacco mosaic virus infection in tobacco plants based on specific RNA hydrolysis induced by a leadzyme. We identified a leadzyme substrate target sequence in genomic tobacco mosaic virus RNA and designed a 16‐mer oligoribonucleotide capable of forming a specific leadzyme motif with a five‐nucleotide catalytic loop. The synthetic 16‐mer RNA was applied with nontoxic, catalytic amount of lead to infected tobacco leaves. We observed inhibition of tobacco mosaic virus infection in tobacco leaves in vivo due to specific tobacco mosaic virus RNA cleavage effected by leadzyme. A significant reduction in tobacco mosaic virus accumulation was observed even when the leadzyme was applied up to 2 h after inoculation of leaves with tobacco mosaic virus. This process, called leadzyme interference, is determined by specific recognition and cleavage of the target site by the RNA catalytic strand in the presence of Pb(2+).