The potent and selective ‘genetic zipper’ method for insect pest control consists of three essential components: an antisense DNA (the finder), its complementary mature rRNA or pre-rRNA of the pest (the target), and the host’s endogenous DNA-guided rRNase (the degrader). Although this approach has been validated, the spectrum of effective rRNA targets remains insufficiently explored. In this study, we report for the first time the insecticidal efficacy of a novel oligonucleotide insecticide, Eriola-11, which targets the mitochondrial 16S rRNA of the woolly apple aphid Eriosoma lanigerum Hausmann. We hypothesized that the antisense-mediated silencing of mitochondrial rRNA would impair aphid viability and lead to physiological disruptions associated with mitochondrial energy metabolism. Eriola-11 was applied either once or twice (with a 24 h interval) to aphid-infested plants, and aphid mortality was recorded over 14 days. Mitochondrial 16S rRNA expression levels were quantified using molecular assays, and the degradation kinetics of Eriola-11 were assessed in aphid tissue homogenates. Results showed significant insecticidal activity, with 67.55% mortality after a single treatment and 83.35% after two treatments. Treated aphids exhibited the loss of their characteristic white woolly wax covering, and mitochondrial 16S rRNA expression was reduced 0.66-fold relative to the control. Additionally, Eriola-11 was fully degraded by aphid DNases from tissue homogenates within 3 h, highlighting its rapid biodegradability. These findings establish mitochondrial 16S rRNA as a viable target for antisense insecticides and expand the catalogue of potential rRNA-based targets, offering a promising avenue for environmentally sustainable pest control strategies.

The article provides basic knowledge on how to calculate selectivity of ‘genetic zipper’ (‘GZ’) tech-nology-based oligonucleotide pesticides to provide safety for non-target organisms. Also here we rep-resent 500 oligonucleotide pesticides computationally predicted and generated in one day via the DNAInsector web tool (dnainsector.com). 11 nucleotides long pesticides targeting pests’ pre-rRNA and rRNA provide high species specificity and theoretical selectivity at the level of 1 in 4,194,304. None of the 500 computationally predicted oligonucleotide pesticides coincide with each other and this essentially shows that for any ecosystem, even if it consists of closely related species, it is possible to create selectively acting oligonucleotide pesticides. ‘GZ’ technology is a smart approach transfer-ring us to new possibilities of development of potent and selective oligonucleotide pesticides based on new principles prompted by nature.