The stability and sequence cleavage preference of dsRNA are key factors differentiating RNAi efficiency between migratory locust and Asian corn borer. (April 2022)

• Record Type: Journal Article
• Authors: Fan, Yunhe; Song, Huifang; Abbas, Mureed; Wang, Yanli; Liu, Xiaojian; Li, Tao; Ma, Enbo; Zhu, Kun Yan; Zhang, Jianzhen
• Is Part Of: Insect biochemistry and molecular biology. Volume 143(2022)
• Publication Date: 2022-04
• Subjects: RNA interference -- Locusta migratoria -- Ostrinia furnacalis -- Tissue culture -- Double-stranded RNA -- RNA sequencing -- Small interfering RNAs; Insect biochemistry -- Periodicals; Insects -- Physiology -- Periodicals; Insects -- Molecular aspects -- Periodicals; Biochemistry -- Periodicals; Insectes -- Biochimie -- Périodiques; Insectes -- Composition -- Périodiques; Insectes -- Physiologie -- Périodiques; Insectes -- Aspect moléculaire -- Périodiques; Biochimie -- Périodiques; Biochemistry; Insect biochemistry; Insects -- Molecular aspects; Insects -- Physiology; Periodicals; 572.8157
• DOI: 10.1016/j.ibmb.2022.103738
• Languages: English
• ISSNs: 0965-1748

Abstract

Abstract: We compared the stability of double-stranded RNA (dsRNA) in each of two body fluids (hemolymph, midgut fluid) and in each of two tissues (integument, midgut), and the uptake of dsRNA in each of two cultured tissues (integument, midgut) between the migratory locust ( Locusta migratoria ) and the Asian corn borer ( Ostrinia furnacalis ). We further compared the abundance of putative small interfering RNAs (siRNAs) generated from each of two dsRNAs (ds β-actin, ds Ef1α ) and the preference of dsRNA cleavages between the two insect species. Our studies showed a rapid degradation of dsRNA in the midgut fluids of both insect species and in O. furnacalis hemolymph. However, dsRNA remained reasonably stable in L. migratoria hemolymph. When nuclease degradation of dsRNA in cultured tissues was inhibited, dsRNA uptake was not significantly different between the two species. We further showed that the silencing efficiency against target genes was consistent with the abundance of putative siRNAs processed from the dsRNA. In addition, O. furnacalis showed a strong preference in cleaving dsRNA when the nucleotide G was in the position of "1" at 5′-end whereas L. migratoria showed broad spectrum in cleavage sites to generate siRNA. Taken together, our study revealed that silencing efficiency of a target gene by RNAi was directly related to the dsRNA degradation by nucleases and the abundance of siRNAs generated from the dsRNA. Graphical abstract: Image 1 Highlights: Factors affecting RNAi efficiency were compared in L. migratoria and O. furnacalis . DsRNA up-take by cultured integument or midgut was similar between the two species. Low RNAi efficiency was related to a rapid dsRNA degradation by nucleases. High RNAi efficiency was related to high siRNA abundance generated from dsRNA. O. furnacalis showed strong dsRNA cleavage preference but L. migratoria did not.