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Insects
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Challenges and Future Trends of RNA Interference in Insects
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Challenges and Future Trends of RNA Interference in Insects

A special issue of Insects (ISSN 2075-4450). This special issue belongs to the section "Insect Molecular Biology and Genomics".

Deadline for manuscript submissions: closed (30 September 2024) | Viewed by 11642

Special Issue Editor

Dr. Honglin Feng

E-Mail Website
Guest Editor
Department of Entomology, Louisiana State University, Baton Rouge, LA 70803, USA
Interests: RNA interference; insects; pest management; RNAi delivery; RNAi safety

Special Issue Information

Dear Colleagues,

RNA interference (RNAi) is a powerful molecular tool that has revolutionized functional genomics and gene regulation in various organisms, including insects. Insects are critical model systems for understanding the mechanisms of RNAi and developing novel RNAi-based technologies for pest management. However, despite the significant progress made in the field, several challenges and future directions need to be addressed to fully exploit RNAi’s potential in insects. The objective of this Special Issue is to provide a comprehensive overview of the current challenges and future trends of RNAi in insects. This Special Issue aims to cover a range of topics, including the mechanisms of RNAi in insects, novel delivery strategies, target gene selection, off-target effects, insect resistance, and the application of RNAi in insect pest management. This Special Issue will bring together leading experts in the field to discuss the latest advancements in RNAi technology, highlight the potential of RNAi as a tool for managing insect pests, and identify key areas for future research.

Dr. Honglin Feng
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Insects is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • RNA interference
  • insects
  • pest management
  • RNAi delivery
  • RNAi safety

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Published Papers (6 papers)

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Research

13 pages, 5787 KiB  
Article
Chitin Deacetylase 1 Gene as an Optimal RNAi-Based Target for Controlling the Tomato Leaf Miner Tuta absoluta
by Yangfan Zhou, Yu Zhang, Kangkang Xu, Ruiyu Liu, Wenbiao Liu, Hang Ma and Wenjia Yang
Insects 2024, 15(11), 838; https://doi.org/10.3390/insects15110838 - 25 Oct 2024
Viewed by 1719
Abstract
Chitin is a critical component of both the exoskeleton and internal structures of insects, which can protect insects from mechanical damage, dehydration and pathogen infection, and plays a significant role in the molting process. Chitin deacetylases (CDAs), key enzymes involved in chitin metabolism, [...] Read more.
Chitin is a critical component of both the exoskeleton and internal structures of insects, which can protect insects from mechanical damage, dehydration and pathogen infection, and plays a significant role in the molting process. Chitin deacetylases (CDAs), key enzymes involved in chitin metabolism, are widely distributed among arthropods and microorganisms. In this study, we identified a CDA gene, TaCDA1, in the invasive insect species Tuta absoluta (Meyrick). Sequence analysis demonstrated a high degree of similarity to CDAs in other insects, revealing the presence of three conserved domains. Quantitative analysis showed that the TaCDA1 gene exhibited peak expression during the pupal stage, particularly within the epidermis. The suppression of TaCDA1 expression through RNA interference in T. absoluta pupae significantly impacted the expression of genes associated with chitin metabolism, increasing mortality and developmental abnormalities during the pupa–adult transition and reducing the pupal weight. Furthermore, soaking gene-specific dsRNA resulted in elevated mortality rates during the larva–pupa transition, causing the inability to form new cuticles or undergo ecdysis, as confirmed by subsequent histological observations. The oral administration of dsTaCDA1 + sucrose solution did not significantly impact NtCDA1 expression or the mortality rate compared to the dsGFP + sucrose solution control in the non-target insect Nesidiocoris tenuis. This study demonstrated that TaCDA1 is a potential and safe target for pest control of T. absoluta. Full article
(This article belongs to the Special Issue Challenges and Future Trends of RNA Interference in Insects)
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12 pages, 2743 KiB  
Article
RNA Interference Reveals the Impacts of CYP6CY7 on Imidacloprid Resistance in Aphis glycines
by Shuangyu Li, Hongjia Yang, Yixiao Wang, Lisi Wei, Jiawei Lyu, Zhimeng Shan, Xinxin Zhang and Dong Fan
Insects 2024, 15(3), 188; https://doi.org/10.3390/insects15030188 - 13 Mar 2024
Cited by 2 | Viewed by 1491
Abstract
Cytochrome P450 (CYP) is a group of important detoxification enzymes found in insects related to their resistance to insecticides. To elucidate the CYP6 family genes of P450, which are potentially related to imidacloprid resistance in Aphis glycines, the CYP6 cDNA sequences of [...] Read more.
Cytochrome P450 (CYP) is a group of important detoxification enzymes found in insects related to their resistance to insecticides. To elucidate the CYP6 family genes of P450, which are potentially related to imidacloprid resistance in Aphis glycines, the CYP6 cDNA sequences of A. glycines were studied. The transcriptome of A. glycines was constructed, and the CYP6 cDNA sequences of A. glycines were screened. Their relative expression levels in response to imidacloprid induction were examined through qRT-PCR, and the CYP6s with higher expression levels were used to study the detoxification of imidacloprid through RNA interference and a bioassay. Twelve CYP6s were obtained from the A. glycines transcriptome. These samples were named by the International P450 Nomenclature Committee and registered in GenBank. After 3, 6, 12, 24 and 48 h of induction with LC50 concentrations of imidacloprid, the relative expression levels of these CYP6s increased; the expression level of CYP6CY7 experienced the highest increase, being more than 3-fold higher than that of those of the non-imidacloprid-induced CYP6s. After RNA interference for CYP6CY7, the relative expression level of CYP6CY7 significantly decreased after 3, 6 and 12 h, while the corresponding P450 enzyme activity decreased after 12 and 24 h. The mortality of A. glycines due to imidacloprid treatment increased by 14.71% at 24 h. CYP6CY7 might detoxify imidacloprid in A. glycines. This study provides a theoretical basis for the further study of the mechanism of action of CYP6s and potential new methods for improving insecticidal efficacy. Full article
(This article belongs to the Special Issue Challenges and Future Trends of RNA Interference in Insects)
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12 pages, 1885 KiB  
Article
Cyantraniliprole and Thiamethoxam Exposure Changes Expression of Transcripts Associated with Small Non-Coding RNA Processing in the Colorado Potato Beetle
by Pierre Bastarache, Kenan Timani, Mariem Ben Youssef, Enock Omakele, Jess L. Vickruck and Pier Jr. Morin
Insects 2024, 15(3), 147; https://doi.org/10.3390/insects15030147 - 22 Feb 2024
Viewed by 1588
Abstract
The Colorado potato beetle (Leptinotarsa decemlineata (Say)) can cause extensive damage to agricultural crops worldwide and is a significant insect pest. This insect is notorious for its ability to evade various strategies deployed to control its spread and is known for its [...] Read more.
The Colorado potato beetle (Leptinotarsa decemlineata (Say)) can cause extensive damage to agricultural crops worldwide and is a significant insect pest. This insect is notorious for its ability to evade various strategies deployed to control its spread and is known for its relative ease in developing resistance against different insecticides. Various molecular levers are leveraged by L. decemlineata for this resistance to occur, and a complete picture of the genes involved in this process is lacking. While small non-coding RNAs, including miRNAs, are differentially expressed in insects exposed to insecticides, levels of transcript coding for proteins underlying their synthesis remain to be characterized fully. The overarching objective of this work aims to fill that gap by assessing the expression of such targets in L. decemlineata exposed to cyantraniliprole and thiamethoxam. The expression status of Ago1, Ago2, Ago3, Dcr2a, Dcr2b, Expo-5, Siwi-1 and Siwi-2 transcripts were quantified via qRT-PCR in adult L. decemlineata treated with low and high doses of these compounds for different lengths of time. Variation in Ago1 and Dcr2b expression was notably observed in L. decemlineata exposed to cyantraniliprole, while thiamethoxam exposure was associated with the modulation of Dcr2a and Siwi-1 transcript levels. The down-regulation of Ago1 expression in L. decemlineata using dsRNA, followed by cyantraniliprole treatment, was associated with a reduction in the survival of insects with reduced Ago1 transcript expression. Overall, this work presents the insecticide-mediated modulation of transcripts associated with small non-coding RNA processing and showcases Ago1 as a target to further investigate its relevance in cyantraniliprole response. Full article
(This article belongs to the Special Issue Challenges and Future Trends of RNA Interference in Insects)
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18 pages, 9389 KiB  
Article
Knockdown of the Expression of Two Trehalase Genes with RNAi Disrupts the Trehalose and Chitin Metabolism Pathways in the Oriental Armyworm, Mythimna separata
by Hongjia Yang, Yixiao Wang, Weijia Zhang, Xinxin Zhang, Sibo Wang, Mengyao Cui, Xiaohui Zhao, Dong Fan and Changchun Dai
Insects 2024, 15(3), 142; https://doi.org/10.3390/insects15030142 - 21 Feb 2024
Cited by 2 | Viewed by 1475
Abstract
Trehalose is an important carbohydrate substance in insect hemolymph. Chitin is the main component of cuticle and peritrophic matrix in insects. Trehalase (Tre) catalyzes the decomposition of trehalose. Few studies of trehalase in lepidopteran insects have been conducted. Here, the functions of soluble [...] Read more.
Trehalose is an important carbohydrate substance in insect hemolymph. Chitin is the main component of cuticle and peritrophic matrix in insects. Trehalase (Tre) catalyzes the decomposition of trehalose. Few studies of trehalase in lepidopteran insects have been conducted. Here, the functions of soluble Tre (Tre1) and membrane-bound Tre (Tre2) in the growth and development of Mythimna separata were investigated. We cloned and identified Tre1 and Tre2 cDNA sequences in M. separata. Analysis expression revealed that MsTre1 and MsTre2 were highly expressed in midgut and integument, respectively. The expression of MsTre1 and MsTre2 was highest in the pupal stage. We used RNA interference (RNAi) to inhibit Tre expression in M. separata larvae. Injection of dsMsTre1 or dsMsTre2 resulted in abnormal phenotypes and impeded normal molting. Silencing of MsTre1 and MsTre2 resulted in significant changes in the expression of genes in the trehalose and chitin metabolism pathways, significantly increased the trehalose and glycogen content, and significantly decreased MsTre1 and MsTre2 activity, the glucose content, and the chitin content in midgut and integument. Silencing of MsTre1 slowed larval molting, and the new cuticle was significantly thinner. These results indicate that RNAi of Tre may be useful for control strategies against M. separata. Full article
(This article belongs to the Special Issue Challenges and Future Trends of RNA Interference in Insects)
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14 pages, 2525 KiB  
Article
RNAi-Mediated Knockdown of Acidic Ribosomal Stalk Protein P1 Arrests Egg Development in Adult Female Yellow Fever Mosquitoes, Aedes aegypti
by Mahesh Lamsal, Hailey A. Luker, Matthew Pinch and Immo A. Hansen
Insects 2024, 15(2), 84; https://doi.org/10.3390/insects15020084 - 24 Jan 2024
Viewed by 2450
Abstract
After taking a blood meal, the fat body of the adult female yellow fever mosquito, Aedes aegypti, switches from a previtellogenic state of arrest to an active state of synthesizing large quantities of yolk protein precursors (YPPs) that are crucial for egg [...] Read more.
After taking a blood meal, the fat body of the adult female yellow fever mosquito, Aedes aegypti, switches from a previtellogenic state of arrest to an active state of synthesizing large quantities of yolk protein precursors (YPPs) that are crucial for egg development. The synthesis of YPPs is regulated at both the transcriptional and translational levels. Previously, we identified the cytoplasmic protein general control nonderepressible 1 (GCN1) as a part of the translational regulatory pathway for YPP synthesis. In the current study, we used the C-terminal end of GCN1 to screen for protein–protein interactions and identified 60S acidic ribosomal protein P1 (P1). An expression analysis and RNAi-mediated knockdown of P1 was performed to further investigate the role of P1 in mosquito reproduction. We showed that in unfed (absence of a blood meal) adult A. aegypti mosquitoes, P1 was expressed ubiquitously in the mosquito organs and tissues tested. We also showed that the RNAi-mediated knockdown of P1 in unfed adult female mosquitoes resulted in a strong, transient knockdown with observable phenotypic changes in ovary length and egg deposition. Our results suggest that 60S acidic ribosomal protein P1 is necessary for mosquito reproduction and is a promising target for mosquito population control. Full article
(This article belongs to the Special Issue Challenges and Future Trends of RNA Interference in Insects)
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12 pages, 1806 KiB  
Article
Demonstration of RNAi Yeast Insecticide Activity in Semi-Field Larvicide and Attractive Targeted Sugar Bait Trials Conducted on Aedes and Culex Mosquitoes
by Akilah T. M. Stewart, Keshava Mysore, Teresia M. Njoroge, Nikhella Winter, Rachel Shui Feng, Satish Singh, Lester D. James, Preeraya Singkhaimuk, Longhua Sun, Azad Mohammed, James D. Oxley, Craig Duckham, Alongkot Ponlawat, David W. Severson and Molly Duman-Scheel
Insects 2023, 14(12), 950; https://doi.org/10.3390/insects14120950 - 15 Dec 2023
Cited by 1 | Viewed by 2296
Abstract
Eco-friendly new mosquito control innovations are critical for the ongoing success of global mosquito control programs. In this study, Sh.463_56.10R, a robust RNA interference (RNAi) yeast insecticide strain that is suitable for scaled fermentation, was evaluated under semi-field conditions. Inactivated and dried Sh.463_56.10R [...] Read more.
Eco-friendly new mosquito control innovations are critical for the ongoing success of global mosquito control programs. In this study, Sh.463_56.10R, a robust RNA interference (RNAi) yeast insecticide strain that is suitable for scaled fermentation, was evaluated under semi-field conditions. Inactivated and dried Sh.463_56.10R yeast induced significant mortality of field strain Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus larvae in semi-field larvicide trials conducted outdoors in St. Augustine, Trinidad, where 100% of the larvae were dead within 24 h. The yeast was also stably suspended in commercial bait and deployed as an active ingredient in miniature attractive targeted sugar bait (ATSB) station sachets. The yeast ATSB induced high levels of Aedes and Culex mosquito morbidity in semi-field trials conducted in Trinidad, West Indies, as well as in Bangkok, Thailand, in which the consumption of the yeast resulted in adult female mosquito death within 48 h, faster than what was observed in laboratory trials. These findings support the pursuit of large-scale field trials to further evaluate the Sh.463_56.10R insecticide, a member of a promising new class of species-specific RNAi insecticides that could help combat insecticide resistance and support effective mosquito control programs worldwide. Full article
(This article belongs to the Special Issue Challenges and Future Trends of RNA Interference in Insects)
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