siRNA enrichment in Argonaute 2-depleted Blattella germanica

Autor: José Carlos Montañés, Carlos Rojano, Maria-Dolors Piulachs, Guillem Ylla, José L. Maestro
Přispěvatelé: Ministerio de Economía y Competitividad (España), Generalitat de Catalunya, Consejo Superior de Investigaciones Científicas (España)
Rok vydání: 2021
Předmět:
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
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ISSN: 1876-4320
2016-7601
Popis: [Background] RNA interference (RNAi) is a cellular mechanism used to fight various threats, including transposons, aberrant RNAs, and some types of viruses. This mechanism relies on the detection of dsRNA molecules, which through a pathway involving Dicer-2 (Dcr-2) and Argonaute 2 (AGO2), produces small interfering RNAs (siRNAs) that bind to the complementary RNAs triggering their degradation.
[Methods] Using the cockroach Blattella germanica as a model, we examined AGO2 activity by depleting its mRNA using RNAi and analyzing the phenotypes produced.
[Results] Depleting AGO2 expression had no remarkable effect on nymphal development or reproduction. dsRNA treatment triggered an immediate and transitory increase in AGO2 expression, independently of Dcr-2 action. In addition, we analyzed the siRNAs generated after injecting a heterologous dsRNA in control and AGO2-depleted animals. The results revealed that obtained siRNAs mapped non-uniformly along the dsRNA sequence. In AGO2-depleted animals, the proportion of 22 nucleotide reads was higher and accumulations of reads appeared in areas less well-represented in the controls. We also detected a preference for cytosine as the first nucleotide in controls that was significantly attenuated in AGO2-depleted individuals.
[Conclusions/general significance] The siRNAs produced from a dsRNA mapped heterogeneously along the length of the dsRNA and this arrangement depends on the dsRNA sequence. AGO2 exerts its role as nuclease on the siRNA duplexes independently of its action on the corresponding mRNA. This study sheds light on an extremely useful process for reverse genetics in laboratories, in addition to the design of more effective, specific, and eco-friendly pest-control strategies.
This work was supported by Spanish Ministry of Economy and Competitiveness and FEDER (grant number CGL2016-76011-R), Secretaria d'Universitats i Recerca, Catalan Goverment (grant number 2017 SGR 1030), Agencia Estatal de Investigación (grant number PID2019-104483GB-I00/AEI/10.13039/501100011033), and Agencia Estatal Consejo Superior de Investigaciones Científicas (grant numbers 2019AEP028 and 2019AEP029).
Databáze: OpenAIRE
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