Double-Strand RNA (dsRNA) Delivery Methods in Insects: Diaphorina citri

Santos‐Ortega, Yulica; Flynt, Alex S.

doi:10.1007/978-1-0716-1633-8_19

Cited by 4 publications

(2 citation statements)

References 132 publications

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“…The microinjection technique offers the precise and direct introduction of a defined quantity and concentration of miRNA (mimics or inhibitors) into the desired location. Nonetheless, it requires substantial labor, specialized knowledge, and equipment rendering it unsuitable for field-based applications [87].…”

Section: Microinjectionmentioning

confidence: 99%

miRNA Dynamics for Pest Management: Implications in Insecticide Resistance

Mahalle,

Mota-Sanchez,

Pittendrigh

et al. 2024

Insects

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Utilizing chemical agents in pest management in modern agricultural practices has been the predominant approach since the advent of synthetic insecticides. However, insecticide resistance is an emerging issue, as pest populations evolve to survive exposure to chemicals that were once effective in controlling them, underlining the need for advanced and innovative approaches to managing pests. In insects, microRNAs (miRNAs) serve as key regulators of a wide range of biological functions, characterized by their dynamic expression patterns and the ability to target genes. Recent studies are increasingly attributed to the significance of miRNAs in contributing to the evolution of insecticide resistance in numerous insect species. Abundant miRNAs have been discovered in insects using RNA sequencing and transcriptome analysis and are known to play vital roles in regulation at both the transcriptional and post-transcriptional levels. Globally, there is growing research interest in the characterization and application of miRNAs, especially for their potential role in managing insecticide resistance. This review focuses on how miRNAs contribute to regulating insecticide resistance across various insect species. Furthermore, we discuss the gain and loss of functions of miRNAs and the techniques for delivering miRNAs into the insect system. The review emphasizes the application of miRNA-based strategies to studying their role in diminishing insecticide resistance, offering a more efficient and lasting approach to insect management.

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Section: Microinjectionmentioning

confidence: 99%

miRNA Dynamics for Pest Management: Implications in Insecticide Resistance

Mahalle,

Mota-Sanchez,

Pittendrigh

et al. 2024

Insects

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“…Microinjection is one of the most efficient and widely used methods to introduce dsRNA in insects (Yang & Han, 2014). Besides various dsRNA delivery methods like soaking and diet incorporation feeding, dsRNA injection has been quite successful in insects including Lepidoptera (Santos‐Ortega & Flynt, 2022; Yu et al, 2013). The success of RNAi experiments in insects is often governed by several factors, of which the precise amount of dsRNA delivery holds paramount importance.…”

Section: Introductionmentioning

confidence: 99%

Simple cost‐effective larval injection method for dsRNA delivery to induce RNAi response in Helicoverpa armigera (Hübner)

Das

Kumar

Shah

et al. 2023

J Applied Entomology

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Microinjection is considered as an effective method for dsRNA delivery in insects. It also facilitates the delivery of a precise quantity of dsRNA in the host insect, inducing an efficient RNAi response. However, the microinjection method needs prior optimization of several parameters like concentration of dsRNA, site of injection, developmental stage of insect etc. for inducing effective RNAi response. Moreover, sophisticated microinjection devices are largely expensive with high maintenance cost. The Old‐World bollworm, Helicoverpa armigera (Hübner) is known to be a detrimental polyphagous pest with widespread infestations across the globe. In the present study, we demonstrate a low‐cost and effective dsRNA delivery method for inducing RNAi response in H. armigera with the aid of basic insulin injection syringe and fabricated micropipette tip. In order to validate the RNAi response following dsRNA injection, we have selected three key genes from the chitin biosynthesis pathway of the insect. Besides these, argonaute 1 (ago1) was also used as an indicator gene for dsRNA‐mediated RNAi induction. Delivery of dsRNA using injection with insulin syringe caused significant upregulation of the ago1 gene in the insect irrespective of any of the three target genes concerned viz. HaNAGK (3.9 fold; p < 0.001), HaAGM (6.3 fold; p < 0.001) and HaUAP (5.9 fold; p < 0.01) respectively, as compared to control injected with nuclease‐free water. The dsRNA‐injected insects showed aberrant developmental symptoms typical of impeded chitin synthesis, eventually leading to anomalous ecdysis with substantial mortality (up to 69.04%), as compared to controls. The described protocol reduces insect injury, enabling easy restraining of larva and quick execution of dsRNA injection with efficient RNAi response.

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