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MiRNA-mediated gene silencing in Drosophila larval development involves GW182-dependent and independent mechanisms

ABSTRACT: microRNAs (miRNAs) regulate a wide variety of biological processes by silencing their target genes. Argonaute (AGO) proteins load miRNAs to form an RNA-induced silencing complex (RISC), which mediates translational repression and/or mRNA decay of the targets. A scaffold protein called GW182 directly binds AGO and the CCR4-NOT deadenylase complex, initiating the mRNA decay reaction. Although previous studies have demonstrated the critical role of GW182 in cultured cells as well as in cell-free systems, its biological significance in living organisms remains poorly explored, especially in Drosophila melanogaster. Here, we generated gw182-null flies using the CRISPR/Cas9 system and found that, unexpectedly, they can survive until an early second-instar larval stage. Moreover, in vivo miRNA reporters can be effectively repressed in gw182-null first-instar larvae. Nevertheless, gw182-null flies have defects in the expression of chitin-related genes and the formation of the larval trachea system, preventing them from completing larval development. Our results highlight the importance of both GW182-dependent and -independent silencing mechanisms in vivo.

SUBMITTER: Dr. Eriko Matsuura-Suzuki

PROVIDER: S-SCDT-10_1038-S44318-024-00249-4 | biostudies-other |

REPOSITORIES: biostudies-other

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miRNA-mediated gene silencing in Drosophila larval development involves GW182-dependent and independent mechanisms

Project description:microRNAs (miRNAs) regulate a wide variety of biological processes by silencing their target genes. Argonaute (AGO) proteins load miRNAs to form RNA-induced silencing complex (RISC), which mediates translational repression and/or mRNA decay of the targets. A scaffold protein called GW182 directly binds AGO and the CCR4-NOT deadenylase complex, initiating the mRNA decay reaction. Although previous studies have demonstrated the critical role of GW182 in vitro and in cultured cells, its biological significance in living organisms remains largely unexplored. Here, we generated gw182-null flies using the CRISPR/Cas9 system and found that, unexpectedly, they can survive until an early second instar larval stage. Moreover, in vivo miRNA reporters can be effectively repressed even in the absence of GW182. Nevertheless, gw182-null flies have defects in expression of chitin-related genes and formation of the larval trachea system, preventing them to complete the larval development. Our results highlight the importance of both GW182-depedent and -independent silencing mechanisms in vivo.

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miRNA-mediated gene silencing in Drosophila larval development involves GW182-dependent and independent mechanisms

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miRNA-mediated gene silencing in Drosophila larval development involves GW182-dependent and independent mechanisms

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miRNA-mediated gene silencing in Drosophila larval development involves GW182-dependent and independent mechanisms

Project description:miRNA-mediated gene silencing in Drosophila larval development involves GW182-dependent and independent mechanisms

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WIG1 is crucial for AGO2-mediated ACOT7 mRNA silencing via miRNA-dependent and -independent mechanisms.

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