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Abstract <p>RNA interference (RNAi) has come up as a promising future technique in the hunt for fresh and alternative approaches to pest management. However, different insect species demonstrate a differential level of response towards RNAi. The first barrier encountered by dsRNA ingested by the insect is the DsRNases specific nucleases in gut and hemolymph. Numerous studies demonstrate poor response of <i>Helicoverpa armigera</i> (<i>H. armigera</i>) to the dsRNA against any target gene. The fate of dsRNA in the haemolymph of <i>H. armigera</i> was assessed through timed incubations of 1, 3, and 5&#xa0;h, which revealed that haemolymph-mediated degradation significantly contributes to the insect’s poor RNAi response.<i>.</i>The degree of degradation of naked dsRNA in hemolymph was directly proportional to the incubation time of dsRNA in hemolymph and concentration of hemolymph. Furthermore, the conjugation of dsRNA with chitosan nanocarrier resulted in a decrease in the impact of haemolymph carried DsRNases on dsRNA and consequently resulting in improved RNAi efficiency. Three genes <i>Aminopeptidase</i> (AMN)<i>, Cadherin</i> (CAD) <i>and Alpha amylase</i> (AMY) were targeted&#xa0;through feeding of chitosan tethered dsRNA showed that feeding chitosan coated dsAMN dramatically decreased the target gene expressionby 76.6% when compared to 13.02% with feeding the same amount naked dsRNA. Similarly, the knockdown efficiency of alpha amylase and cadherin genes was enhanced to&#xa0;91.6 and 76.6%, respectively by tethering the dsRNA with chitosan and delivering it to the insect orally. The simultaneous knockdown of the <i>H. armigera dsRNaes</i> and the <i>Cadherin</i> gene with both the Chitosan-dsRNA nanocarrier resulted in increasing the knockdown efficiency two times compared to&#xa0;administration of naked dsDSRNases and dsCadherin. These studies elucidate the role of insect gut fluids in hampering the RNAi efficiency in the lepidopteran insect <i>H. armigera</i> and its enhancement through Chitosan- dsRNA nanocarrier.</p> Graphical abstract <p></p>

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Overcoming gut and hemolymph barriers: Chitosan nanocarrier enhance dsRNA stability and RNAi in Helicoverpa armigera

  • 期刊论文

  • 发表日期：

    2025年12月14日

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  • Shelja Sandal,
  • Gulshan Bansal,
  • Ramandeep Kaur,
  • Satnam Singh

摘要

Abstract

RNA interference (RNAi) has come up as a promising future technique in the hunt for fresh and alternative approaches to pest management. However, different insect species demonstrate a differential level of response towards RNAi. The first barrier encountered by dsRNA ingested by the insect is the DsRNases specific nucleases in gut and hemolymph. Numerous studies demonstrate poor response of Helicoverpa armigera (H. armigera) to the dsRNA against any target gene. The fate of dsRNA in the haemolymph of H. armigera was assessed through timed incubations of 1, 3, and 5 h, which revealed that haemolymph-mediated degradation significantly contributes to the insect’s poor RNAi response..The degree of degradation of naked dsRNA in hemolymph was directly proportional to the incubation time of dsRNA in hemolymph and concentration of hemolymph. Furthermore, the conjugation of dsRNA with chitosan nanocarrier resulted in a decrease in the impact of haemolymph carried DsRNases on dsRNA and consequently resulting in improved RNAi efficiency. Three genes Aminopeptidase (AMN), Cadherin (CAD) and Alpha amylase (AMY) were targeted through feeding of chitosan tethered dsRNA showed that feeding chitosan coated dsAMN dramatically decreased the target gene expressionby 76.6% when compared to 13.02% with feeding the same amount naked dsRNA. Similarly, the knockdown efficiency of alpha amylase and cadherin genes was enhanced to 91.6 and 76.6%, respectively by tethering the dsRNA with chitosan and delivering it to the insect orally. The simultaneous knockdown of the H. armigera dsRNaes and the Cadherin gene with both the Chitosan-dsRNA nanocarrier resulted in increasing the knockdown efficiency two times compared to administration of naked dsDSRNases and dsCadherin. These studies elucidate the role of insect gut fluids in hampering the RNAi efficiency in the lepidopteran insect H. armigera and its enhancement through Chitosan- dsRNA nanocarrier.

Graphical abstract