Insecticidal activity of three 10–12 nucleotides long antisense sequences from 5.8S ribosomal RNA gene of gypsy moth Lymantria dispar L. against its larvae
Insecticidal activity of three 10–12 nucleotides long antisense sequences from 5.8S ribosomal RNA gene of gypsy moth Lymantria dispar L. against its larvae
5.8S ribosomal RNA plays an important role in protein synthesis and eukaryotic ribosome
translocation. Contact DNA insecticides based on antisense fragments of 5.8S ribosomal
RNA gene of gypsy moth Lymantria dispar L. showed prospective insecticidal activity on its
larvae. The most pronounced insecticidal effect was found for antisense fragments 10 and
11 nucleotides long (oligoRIBO-10 and oligoRIBO-11), whereas 12 nucleotides long fragment (oligoRIBO-12) caused the lowest level of insect mortality. This data corresponds to
results obtained earlier using rabbit reticulocyte and wheat germ extracts, where maximum
inhibition of protein synthesis was observed when a relevant oligomer 10-11 nucleotides long
was used, whilst longer chain lengths resulted in reduced inhibition. Using oligoRIBO-11
fragment we have shown penetration of antisense oligonucleotides to insect cells through
insects’ exoskeletons. MALDI technique registered the penetration of the oligoRIBO-11 fragment into insect cells after 30 min and a significant response of insect cells to the applied
oligonucleotide after 60 min, which indicates not only that the oligonucleotide enters the insect cells, but also the synthesis of new substances in response to the applied DNA fragment.
Contact DNA insecticides developed from the L. dispar 5.8S ribosomal RNA gene provide
a novel biotechnology for plant protection using unmodified antisense oligonucleotides.
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