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Characterization, expression patterns, and transcriptional responses of three core RNA interference pathway genes from Ostrinia nubilalis.
J Insect Physiol. 2021 Feb-Mar; 129:104181.JI

Abstract

RNA interference (RNAi) is commonly used in the laboratory to analyze gene function, and RNAi-based pest management strategies are now being employed. Unfortunately, RNAi is hindered by inefficient and highly-variable results when different insects are targeted, especially lepidopterans, such as the European corn borer (ECB), Ostrinia nubilalis (Lepidoptera: Crambidae). Previous efforts to achieve RNAi-mediated gene suppression in ECB revealed low RNAi efficiency with both double-stranded RNA (dsRNA) injection and ingestion. One mechanism that can affect RNAi efficiency in insects is the expression and function of core RNAi pathway genes, such as those encoding Argonaut 2 (Ago2), Dicer 2 (Dcr2), and a dsRNA binding protein (R2D2). To determine if deficiencies in these core RNAi pathway genes contribute to low RNAi efficiency in ECB, full-length complementary DNAs encoding OnAgo2, OnDcr2, and OnR2D2 were cloned, sequenced, and characterized. A comparison of domain architecture suggested that all three predicted proteins contained the necessary domains to function. However, a comparison of evolutionary distances revealed potentially important variations in the first RNase III domain of OnDcr2, the double-stranded RNA binding domains of OnR2D2, and both the PAZ and PIWI domains of OnAgo2, which may indicate functional differences in enzymatic activity between species. Expression analysis indicated that transcripts for all three genes were expressed in all developmental stages and tissues investigated. Interestingly, the introduction of non-target dsRNA into ECB second-instar larvae via microinjection did not affect OnAgo2, OnDcr2, or OnR2D2 expression. In contrast, ingestion of the same dsRNAs resulted in upregulation of OnDcr2 but downregulation of OnR2D2. The unexpected transcriptional responses of the core machinery and the divergence in amino-acid sequence between specific domains in each core RNAi protein may possibly contribute to low RNAi efficiency in ECB. Understanding the contributions of different RNAi pathway components is critical to adapting this technology for use in controlling lepidopteran pests that exhibit low RNAi efficiency.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Cooper AMW
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA. Electronic address: anacooper@ksu.edu.
Song H
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA; Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi 030006, China.
Shi X
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA; Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi 030006, China.
Yu Z
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA.
Lorenzen M
Department of Entomology and Plant Pathology, Campus Box 7613, North Carolina State University, Raleigh, NC 27695, USA.
Silver K
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA.
Zhang J
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA; Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi 030006, China.
Zhu KY
Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USA. Electronic address: kzhu@ksu.edu.

MeSH

AnimalsArgonaute ProteinsGenes, InsectInsect ControlMothsRNA HelicasesRNA InterferenceRNA, Small InterferingRNA-Binding ProteinsRNAi Therapeutics

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

33359365

Citation

Cooper, Anastasia M W., et al. "Characterization, Expression Patterns, and Transcriptional Responses of Three Core RNA Interference Pathway Genes From Ostrinia Nubilalis." Journal of Insect Physiology, vol. 129, 2021, p. 104181.
Cooper AMW, Song H, Shi X, et al. Characterization, expression patterns, and transcriptional responses of three core RNA interference pathway genes from Ostrinia nubilalis. J Insect Physiol. 2021;129:104181.
Cooper, A. M. W., Song, H., Shi, X., Yu, Z., Lorenzen, M., Silver, K., Zhang, J., & Zhu, K. Y. (2021). Characterization, expression patterns, and transcriptional responses of three core RNA interference pathway genes from Ostrinia nubilalis. Journal of Insect Physiology, 129, 104181. https://doi.org/10.1016/j.jinsphys.2020.104181
Cooper AMW, et al. Characterization, Expression Patterns, and Transcriptional Responses of Three Core RNA Interference Pathway Genes From Ostrinia Nubilalis. J Insect Physiol. 2021 Feb-Mar;129:104181. PubMed PMID: 33359365.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization, expression patterns, and transcriptional responses of three core RNA interference pathway genes from Ostrinia nubilalis. AU - Cooper,Anastasia M W, AU - Song,Huifang, AU - Shi,Xuekai, AU - Yu,Zhitao, AU - Lorenzen,Marcé, AU - Silver,Kristopher, AU - Zhang,Jianzhen, AU - Zhu,Kun Yan, Y1 - 2020/12/25/ PY - 2020/10/28/received PY - 2020/12/17/revised PY - 2020/12/22/accepted PY - 2020/12/29/pubmed PY - 2021/9/21/medline PY - 2020/12/28/entrez KW - Argonaute 2 KW - Diabrotica virgifera virgifera KW - Dicer 2 KW - European corn borer KW - R2D2 KW - RNAi efficiency KW - Western corn rootworm SP - 104181 EP - 104181 JF - Journal of insect physiology JO - J Insect Physiol VL - 129 N2 - RNA interference (RNAi) is commonly used in the laboratory to analyze gene function, and RNAi-based pest management strategies are now being employed. Unfortunately, RNAi is hindered by inefficient and highly-variable results when different insects are targeted, especially lepidopterans, such as the European corn borer (ECB), Ostrinia nubilalis (Lepidoptera: Crambidae). Previous efforts to achieve RNAi-mediated gene suppression in ECB revealed low RNAi efficiency with both double-stranded RNA (dsRNA) injection and ingestion. One mechanism that can affect RNAi efficiency in insects is the expression and function of core RNAi pathway genes, such as those encoding Argonaut 2 (Ago2), Dicer 2 (Dcr2), and a dsRNA binding protein (R2D2). To determine if deficiencies in these core RNAi pathway genes contribute to low RNAi efficiency in ECB, full-length complementary DNAs encoding OnAgo2, OnDcr2, and OnR2D2 were cloned, sequenced, and characterized. A comparison of domain architecture suggested that all three predicted proteins contained the necessary domains to function. However, a comparison of evolutionary distances revealed potentially important variations in the first RNase III domain of OnDcr2, the double-stranded RNA binding domains of OnR2D2, and both the PAZ and PIWI domains of OnAgo2, which may indicate functional differences in enzymatic activity between species. Expression analysis indicated that transcripts for all three genes were expressed in all developmental stages and tissues investigated. Interestingly, the introduction of non-target dsRNA into ECB second-instar larvae via microinjection did not affect OnAgo2, OnDcr2, or OnR2D2 expression. In contrast, ingestion of the same dsRNAs resulted in upregulation of OnDcr2 but downregulation of OnR2D2. The unexpected transcriptional responses of the core machinery and the divergence in amino-acid sequence between specific domains in each core RNAi protein may possibly contribute to low RNAi efficiency in ECB. Understanding the contributions of different RNAi pathway components is critical to adapting this technology for use in controlling lepidopteran pests that exhibit low RNAi efficiency. SN - 1879-1611 UR - https://www.unboundmedicine.com/medline/citation/33359365/Characterization_expression_patterns_and_transcriptional_responses_of_three_core_RNA_interference_pathway_genes_from_Ostrinia_nubilalis_ DB - PRIME DP - Unbound Medicine ER -