RNA interference (RNAi) is a form of gene silencing triggered by double-stranded RNA (dsRNA) that operates in all eukaryotic cells. RNAi has been widely investigated in insects to determine the underlying molecular mechanism, to investigate its role in systemic antiviral defense, and to develop strategies for pest control. When insect cells are infected by viruses, viral dsRNA signatures trigger a local RNAi response to block viral replication and generate virus-derived DNA that confers systemic immunity. RNAi-based insect pest control involves the application of exogenous dsRNA targeting genes essential for insect development or survival, but the efficacy of this approach has limited potency in many pests through a combination of rapid dsRNA degradation, inefficient dsRNA uptake/processing, and ineffective RNAi machinery. This could be addressed by dsRNA screening and evaluation, focusing on dsRNA design and off-target management, as well as dsRNA production and delivery. This review summarizes recent progress to determine the role of RNAi in antiviral defense and as a pest control strategy in insects, addressing gaps between our fundamental understanding of the RNAi mechanism and the exploitation of RNAi-based pest control strategies.
| [1] |
An,X., Zhang,W., Ye,C., Smagghe, G., Wang,J.J. and Niu,J.Z. (2022) Discovery of a widespread presence bunyavirus that may have symbiont-like relationships with different species of aphids. Insect Science, 29, 1120–1134.
|
| [2] |
Arraes,F.B.M., Martins-de-Sa, D., Noriega Vasquez,D.D., Melo,B.P., Faheem, M., de Macedo,L.L.P. et al. (2021) Dissecting protein domain variability in the core RNA interference machinery of five insect orders. RNA Biology, 18, 1653–1681.
|
| [3] |
Baum,J.A., Bogaert, T., Clinton,W., Heck,G.R., Feldmann, P., Ilagan,O. et al. (2007) Control of coleopteran insect pests through RNA interference. Nature Biotechnology, 25, 1322–1326.
|
| [4] |
Bellés,X. (2010) Beyond Drosophila: RNAi in vivo and functional genomics in insects. Annual Review of Entomology, 55, 111–128.
|
| [5] |
Bento,F.M., Marques, R.N., Campana,F.B., Demétrio,C.G., Leandro, R.A., Parra,J.R.P. et al. (2020) Gene silencing by RNAi via oral delivery of dsRNA by bacteria in the South American tomato pinworm, Tuta Absoluta. Pest Management Science, 76, 287–295.
|
| [6] |
Bonning,B.C. and Saleh, M.-C. (2021) The interplay between viruses and RNAi pathways in insects. Annual Review of Entomology, 66, 61–79.
|
| [7] |
Caccia,S., Astarita, F., Barra,E., Di Lelio,I., Varricchio, P. and Pennacchio,F. (2020) Enhancement of Bacillus thuringiensis toxicity by feeding Spodoptera littoralis larvae with bacteria expressing immune suppressive dsRNA. Journal of Pest Science, 93, 303–314.
|
| [8] |
Cappelle,K., De Oliveira, C.F.R., Van Eynde,B., Christiaens,O. and Smagghe, G. (2016) The involvement of clathrin-mediated endocytosis and two Sid-1-like transmembrane proteins in double-stranded RNA uptake in the Colorado potato beetle midgut. Insect Molecular Biology, 25, 315–323.
|
| [9] |
Chen,J., Peng,Y., Zhang,H., Wang, K., Zhao,C., Zhu,G. et al. (2021a) Off-target effects of RNAi correlate with the mismatch rate between dsRNA and non-target mRNA. RNA Biology, 18, 1747–1759.
|
| [10] |
Chen,X., Koo,J., Gurusamy,D., Mogilicherla, K. and Reddy Palli,S. (2021b) Caenorhabditis elegans systemic RNA interference defective protein 1 enhances RNAi efficiency in a lepidopteran insect, the fall armyworm, in a tissue-specific manner. RNA Biology, 18, 1291–1299.
|
| [11] |
Darlington,M., Reinders, J.D., Sethi,A., Lu,A.L., Ramaseshadri, P., Fischer,J.R. et al. (2022) RNAi for Western corn rootworm management: lessons learned, challenges, and future directions. Insects, 13, 57.
|
| [12] |
Dong,Y., Wu,M., Zhang,Q., Fu, J., Loiacono,F.V., Yang,Y. et al. (2022) Control of a sap-sucking insect pest by plastid-mediated RNA interference. Molecular Plant, 15, 1176–1191.
|
| [13] |
Fan,Y.H., Song,H.F., Abbas,M., Wang, Y.L., Li,T., Ma,E.B. et al. (2021) A dsRNA-degrading nuclease (dsRNase2) limits RNAi efficiency in the Asian corn borer (Ostrinia furnacalis). Insect Science, 28, 1677–1689.
|
| [14] |
Fan,Y., Abbas,M., Liu,X., Wang, Y., Song,H., Li,T. et al. (2022a) Increased RNAi efficiency by dsEGFP-induced up-regulation of two core RNAi pathway genes (OfDicer2 and OfAgo2) in the Asian corn borer (Ostrinia furnacalis). Insects, 13, 274.
|
| [15] |
Fan,Y., Song,H., Abbas,M., Wang, Y., Liu,X., Li,T. et al. (2022b) The stability and sequence cleavage preference of dsRNA are key factors differentiating RNAi efficiency between migratory locust and Asian corn borer. Insect Biochemistry and Molecular Biology, 143, 103738.
|
| [16] |
Fire,A., Xu,S., Montgomery,M.K., Kostas,S.A., Driver, S.E. and Mello,C.C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391, 806–811.
|
| [17] |
Flockhart,I., Booker, M., Kiger,A., Boutros,M., Armknecht, S., Ramadan,N. et al. (2006) FlyRNAi: the Drosophila RNAi screening center database. Nucleic Acids Research, 34, D489–D494.
|
| [18] |
Fu,J., Xu,S., Lu,H., Li, F., Li,S., Chang,L. et al. (2022) Resistance to RNA interference by plant-derived double-stranded RNAs but not plant-derived short interfering RNAs in Helicoverpa armigera. Plant, Cell & Environment, 45, 1930–1941.
|
| [19] |
Gao,L., Wang,Y., Fan,Y., Abbas, M., Ma,E., Cooper,A.M.W. et al. (2020) Multiple Argonaute family genes contribute to the siRNA-mediated RNAi pathway in Locusta migratoria. Pesticide Biochemistry and Physiology, 170, 104700.
|
| [20] |
Goic,B., Vodovar, N., Mondotte,J.A., Monot,C., Frangeul, L., Blanc,H. et al. (2013) RNA-mediated interference and reverse transcription control the persistence of RNA viruses in the insect model Drosophila. Nature Immunology, 14, 396–403.
|
| [21] |
Gong,C., Yang,Z., Hu,Y., Wu, Q., Wang,S., Guo,Z. et al. (2022) Silencing of the BtTPS genes by transgenic plant-mediated RNAi to control Bemisia tabaci MED. Pest Management Science, 78, 1128–1137.
|
| [22] |
Gurusamy,D., Mogilicherla, K., Shukla,J.N. and Palli,S.R. (2020) Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda. Archives of Insect Biochemistry and Physiology, 104, e21678.
|
| [23] |
Han,S.H., Kim,J.H., Kim,K. and Lee, S.H. (2019) Selection of lethal genes for ingestion RNA interference against western flower thrips, Frankliniella occidentalis, via leaf discmediated dsRNA delivery. Pesticide Biochemistry and Physiology, 161, 47–53.
|
| [24] |
Horn,T., Narov,K.D. and Panfilio,K.A. (2022) Persistent parental RNAi in the beetle Tribolium castaneum involves maternal transmission of long double-stranded RNA. Advanced Genetics, 3, 2100064.
|
| [25] |
Huang,H.J., Ye,Z.X., Wang,X., Yan, X.T., Zhang,Y., He,Y.J. et al. (2021) Diversity and infectivity of the RNA virome among different cryptic species of an agriculturally important insect vector: whitefly Bemisia tabaci. NPJ Biofilms and Microbiomes, 7, 43.
|
| [26] |
Jain,R.G., Fletcher, S.J., Manzie,N., Robinson,K.E., Li,P., Lu,E. et al. (2022) Foliar application of clay-delivered RNA interference for whitefly control. Nature Plants, 8, 535–548.
|
| [27] |
Khajuria,C., Ivashuta, S., Wiggins,E., Flagel,L., Moar,W., Pleau,M. et al. (2018) Development and characterization of the first dsRNA-resistant insect population from western corn rootworm, Diabrotica virgifera virgifera LeConte. PLoS ONE, 13, e0197059.
|
| [28] |
Knorr,E., Fishilevich, E., Tenbusch,L., Frey,M.L.F., Rangasamy, M., Billion,A. et al. (2018) Gene silencing in Tribolium castaneum as a tool for the targeted identification of candidate RNAi targets in crop pests. Scientific Reports, 8, 2061.
|
| [29] |
Li,H., Li,W.X. and Ding,S.W. (2002) Induction and suppression of RNA silencing by an animal virus. Science, 296, 1319–1321.
|
| [30] |
Luo,Y., Chen,Q., Luan,J., Chung, S.H., Van Eck,J., Turgeon,R. et al. (2017) Towards an understanding of the molecular basis of effective RNAi against a global insect pest, the whitefly Bemisia tabaci. Insect Biochemistry and Molecular Biology, 88, 21–29.
|
| [31] |
Luo,Y., Wang,X., Yu,D. and Kang, L. (2012) The SID-1 double-stranded RNA transporter is not required for systemic RNAi in the migratory locust. RNA Biology, 9, 663–671.
|
| [32] |
Mao,Y.B., Cai,W.J., Wang,J.W., Hong, G.J., Tao,X.Y., Wang,L.J. et al. (2007) Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology, 25, 1307–1313.
|
| [33] |
Martinez,Z., De Schutter, K., Van Damme,E.J.M., Vogel,E., Wynant, N., Vanden Broeck,J. et al. (2021) Accelerated delivery of dsRNA in lepidopteran midgut cells by a Galanthus nivalis lectin (GNA)-dsRNA-binding domain fusion protein. Pesticide Biochemistry and Physiology, 175, 104853.
|
| [34] |
Mingels,L., Wynant, N., Santos,D., Peeters,P., Gansemans, Y., Billen,J. et al. (2020) Extracellular vesicles spread the RNA interference signal of Tribolium castaneum TcA cells. Insect Biochemistry and Molecular Biology, 122, 103377.
|
| [35] |
Mishra,S., Dee,J., Moar,W., Dufner-Beattie, J., Baum,J., Dias,N.P. et al. (2021) Selection for high levels of resistance to double-stranded RNA (dsRNA) in Colorado potato beetle (Leptinotarsa decemlineata Say) using non-transgenic foliar delivery. Scientific Reports, 11, 6523.
|
| [36] |
Niu,J., Smagghe, G., De Coninck,D.I.M., Van Nieuwerburgh,F., Deforce, D. and Meeus,I. (2016) In vivo study of Dicer-2-mediated immune response of the small interfering RNA pathway upon systemic infections of virulent and avirulent viruses in Bombus terrestris. Insect Biochemistry and Molecular Biology, 70, 127–137.
|
| [37] |
Niu,J., Taning, C.N.T., Christiaens,O., Smagghe,G. and Wang, J.J. (2018) Rethink RNAi in insect pest control: challenges and perspectives. Advance in Insect Physiology, 55, 1–17.
|
| [38] |
Nwokeoji,A.O., Kumar,S., Kilby,P.M., Portwood, D.E., Hobbs,J.K. and Dickman,M.J. (2019) Analysis of long dsRNA produced in vitro and in vivo using atomic force microscopy in conjunction with ion-pair reverse-phase HPLC. Analyst, 144, 4985–4994.
|
| [39] |
Paradkar,P.N., Trinidad, L., Voysey,R., Duchemin,J.-B. and Walker, P.J. (2012) Secreted Vago restricts West Nile virus infection in Culex mosquito cells by activating the Jak-STAT pathway. Proceedings of the National Academy of Sciences USA, 109, 18915–18920.
|
| [40] |
Peng,Y., Zhu,G.H., Wang,K., Chen, J., Liu,X., Wu,M. et al. (2021) Knockout of SldsRNase1 and SldsRNase2 revealed their function in dsRNA degradation and contribution to RNAi efficiency in the tobacco cutworm, Spodoptera litura. Journal of Pest Science, 94, 1449–1460.
|
| [41] |
Poirier,E.Z., Goic,B., Tomé-Poderti,L., Frangeul,L., Boussier, J., Gausson,V. et al. (2018) Dicer-2-dependent generation of viral DNA from defective genomes of RNA viruses modulates antiviral immunity in insects. Cell Host & Microbe, 23, 353–365.e8.
|
| [42] |
Raje,K.R., Peterson, B.F. and Scharf,M.E. (2018) Screening of 57 candidate double-dtranded RNAs for insecticidal activity against the pest termite Reticulitermes flavipes (Isoptera: Rhinotermitidae). Journal of Economic Entomology, 111, 2782–2787.
|
| [43] |
Rodrigues,T.B., Mishra, S.K., Sridharan,K., Barnes,E.R., Alyokhin, A., Tuttle,R. et al. (2021) First sprayable double-stranded RNA-based biopesticide product targets proteasome subunit beta type-5 in Colorado potato beetle (Leptinotarsa decemlineata). Frontiers in Plant Science, 12, 728652.
|
| [44] |
Saleh,M.C., Tassetto, M., Van Rij,R.P., Goic,B., Gausson, V., Berry,B. et al. (2009) Antiviral immunity in Drosophila requires systemic RNA interference spread. Nature, 458, 346–350.
|
| [45] |
Santos,D., Mingels, L., Vogel,E., Wang,L., Christiaens, O., Cappelle,K. et al. (2019) Generation of virus- and dsRNA-derived siRNAs with species-dependent length in insects. Viruses, 11, 738.
|
| [46] |
Shi,M., Lin,X.D., Tian,J.H., Chen, L.J., Chen,X., Li,C.X. et al. (2016) Redefining the invertebrate RNA virosphere. Nature, 540, 539–543.
|
| [47] |
Spit,J., Philips, A., Wynant,N., Santos,D., Plaetinck, G. and Vanden Broeck,J. (2017) Knockdown of nuclease activity in the gut enhances RNAi efficiency in the Colorado potato beetle, Leptinotarsa decemlineata, but not in the desert locust, Schistocerca gregaria. Insect Biochemistry and Molecular Biology, 81, 103–116.
|
| [48] |
Swevers,L., Liu,J. and Smagghe,G. (2018) Defense mechanisms against viral infection in Drosophila: RNAi and non-RNAi. Viruses, 10, 230.
|
| [49] |
Taning,C.N.T., Gui,S., De Schutter,K., Jahani,M., Castellanos, N.L., Christiaens,O. et al. (2021) A sequence complementarity-based approach for evaluating off-target transcript knockdown in Bombus terrestris, following ingestion of pest-specific dsRNA. Journal of Pest Science, 94, 487–503.
|
| [50] |
Taochy,C., Gursanscky, N.R., Cao,J., Fletcher,S.J., Dressel, U., Mitter,N. et al. (2017) A genetic screen for impaired systemic RNAi highlights the crucial role of DICER-LIKE 2. Plant Physiology, 175, 1424–1437.
|
| [51] |
Tassetto,M., Kunitomi, M. and Andino,R. (2017) Circulating immune cells mediate a systemic RNAi-based adaptive antiviral response in Drosophila. Cell, 169, 314–325.
|
| [52] |
Tassetto,M., Kunitomi, M., Whitfield,Z.J., Dolan,P.T., Sánchez-Vargas, I., Garcia-Knight,M. et al. (2019) Control of RNA viruses in mosquito cells through the acquisition of vDNA and endogenous viral elements. eLife, 8, e41244.
|
| [53] |
Tomari,Y., Du,T., Haley,B., Schwarz, D.S., Bennett,R., Cook,H.A. et al. (2004) RISC assembly defects in the Drosophila RNAi mutant armitage. Cell, 116, 831–841.
|
| [54] |
Ulrich,J., Dao,V.A., Majumdar,U., Schmitt-Engel, C., Schwirz,J., Schultheis,D. et al. (2015) Large scale RNAi screen in Tribolium reveals novel target genes for pest control and the proteasome as prime target. BMC Genomics, 16, 674.
|
| [55] |
Wang,Y., Gong,Q., Wu,Y., Huang, F., Ismayil,A., Zhang,D. et al. (2021) A calmodulin-binding transcription factor links calcium signaling to antiviral RNAi defense in plants. Cell Host & Microbe, 29, 1393–1406.e7.
|
| [56] |
Wang,Z.G., Chen,R.Y., Jiang,Y.K., Wang, Z.W., Wang,J.J. and Niu,J. (2023) Investigation of potential non-target effects to a ladybeetle Propylea japonica in the scenario of RNAi-based pea aphid control. Entomologia Generalis, 43, 79–88.
|
| [57] |
Wu,H., Pang,R., Cheng,T., Xue, L., Zeng,H., Lei,T. et al. (2020) Abundant and diverse RNA viruses in insects revealed by RNA-Seq analysis: ecological and evolutionary implications. Msystems, 5, e00039–e00020.
|
| [58] |
Wu,M., Dong,Y., Zhang,Q., Li, S., Chang,L., Loiacono,F.V. et al. (2022) Efficient control of western flower thrips by plastid-mediated RNA interference. Proceedings of the National Academy of Sciences USA, 119, e2120081119.
|
| [59] |
Xiao,D., Gao,X., Xu,J., Liang, X., Li,Q., Yao,J. et al. (2015) Clathrin-dependent endocytosis plays a predominant role in cellular uptake of double-stranded RNA in the red flour beetle. Insect Biochemistry and Molecular Biology, 60, 68–77.
|
| [60] |
Xu,L., Xu,S., Sun,L., Zhang, Y., Luo,J., Bock,R. et al. (2021) Synergistic action of the gut microbiota in environmental RNA interference in a leaf beetle. Microbiome, 9, 98.
|
| [61] |
Yan,Y., Ham,B.K., Chong,Y.H., Yeh, S.D. and Lucas,W.J. (2020) A plant SMALL RNA-BINDING PROTEIN 1 family mediates cell-to-cell trafficking of RNAi signals. Molecular Plant, 13, 321–335.
|
| [62] |
Ye,C., An,X., Jiang,Y.D., Ding, B.Y., Shang,F., Christiaens,O. et al. (2019) Induction of RNAi core machinery's gene expression by exogenous dsRNA and the effects of pre-exposure to dsRNA on the gene silencing efficiency in the pea aphid (Acyrthosiphon pisum). Frontiers in Physiology, 9, 1906.
|
| [63] |
Ye,C., An,X., Xie,B.Q., Ding, B.Y., Niu,J. and Wang,J.J. (2023) The involvement of systemic RNA interference deficient-1-like (SIL1) in cellular dsRNA uptake in Acyrthosiphon pisum. Insect Science,
|
| [64] |
Ye,C., Hu,X.S., Wang,Z.W., Wei, D., Smagghe,G., Christiaens,O. et al. (2021) Involvement of clathrin-dependent endocytosis in cellular dsRNA uptake in aphids. Insect Biochemistry and Molecular Biology, 132, 103557.
|
| [65] |
Yoon,J.S., Kim,K. and Palli,S.R. (2020) Double-stranded RNA in exosomes: potential systemic RNA interference pathway in the Colorado potato beetle, Leptinotarsa decemlineata. Journal of Asia-Pacific Entomology, 23, 1160–1164.
|
| [66] |
You,L., Zhang,F., Huang,S., Merchant, A., Zhou,X. and Li,Z. (2020) Over-expression of RNA interference (RNAi) core machinery improves susceptibility to RNAi in silkworm larvae. Insect Molecular Biology, 29, 353–362.
|
| [67] |
Zamore,P.D., Tuschl, T., Sharp,P.A. and Bartel,D.P. (2000) RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell, 101, 25–33.
|
| [68] |
Zhang,H., Gao,J., Chen,J., Peng, Y. and Han,Z. (2022a) RNA-dependent RNA polymerase could extend the lasting validity period of exogenous dsRNA. Pest Management Science, 78, 4569–4578.
|
| [69] |
Zhang,J., Ye,C., Wang,Z.G., Ding, B.Y., Smagghe,G., Zhang,Y. et al. (2022b) dsRNAs spray enhanced the virulence of entomopathogenic fungi Beauveria bassiana in aphid control. Journal of Pest Science, 96, 241–251.
|
| [70] |
Zhang,W., Zhang,Y.C., Wang,Z.G., Gu, Q.Y., Niu,J.Z. and Wang,J.J. (2021) The diversity of viral community in invasive fruit flies (Bactrocera and Zeugodacus) revealed by meta-transcriptomics. Microbial Ecology, 83, 739–752.
|
| [71] |
Zhang,Y.H., Ma,Z.Z., Zhou,H., Chao, Z.J., Yan,S. and Shen,J. (2022c) Nanocarrier-delivered dsRNA suppresses wing development of green peach aphids. Insect Science, 29, 669–682.
|
| [72] |
Zhao,J. and Lei, T. (2020) A vector whitefly endocytic receptor facilitates the entry of begomoviruses into its midgut cells via binding to virion capsid proteins. PLoS Pathogens, 16, e1009053.
|
| [73] |
Zhu,K.Y. and Palli, S.R. (2020) Mechanisms, applications, and challenges of insect RNA interference. Annual Review of Entomology, 65, 293–311.
|
RIGHTS & PERMISSIONS
2023 The Authors. Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.
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