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Published VersionCC BY V4.0, Open Access
Journal article
Silencing Multiple Crustacean Hyperglycaemic Hormone-Encoding Genes in the Redclaw Crayfish Cherax quadricarinatus Induces Faster Molt Rates with Anomalies
International Journal of Molecular Sciences, Vol.25(22), pp.1-18
2024
Abstract
RNA interference (RNAi)-based biotechnology has been previously implemented in decapod crustaceans. Unlike traditional RNAi methodologies that investigate single gene silencing, we employed a multigene silencing approach in decapods based on chimeric double-stranded RNA (dsRNA) molecules coined ‘gene blocks’. Two dsRNA constructs, each targeting three genes of the crustacean hyperglycaemic hormone (CHH) superfamily of neuropeptides, were produced: Type II construct targeting Cq-Molt-inhibiting hormone 1 (MIH1), Cq-MIH-like 1 (MIHL1), and Cq-MIHL2 isoforms and Type I construct targeting Cq-ion transport peptide (Cq-ITP; a putative hybrid of CHH and MIH) and Cq-CHH and Cq-CHH-like (CHHL) isoforms. Both constructs were injected into juvenile redclaw crayfish, Cherax quadricarinatus, to determine the effects of multigene knockdown on molting and developmental processes. A 20-Hydroxyecdysone (20E) enzyme-linked immunosorbent assay (ELISA) and glucose assay were used to determine the effects of RNAi on molting and hemolymph glycemic activities, respectively. Multigene silencing reduced the intermolt interval by 23%. Statistically significant elevated 20E was recorded in treated intermolt individuals, consistent with the reduced intermolt interval as well as unique and abnormal phenotypes related to the molting process, which indicates a shift in 20E-induced cascade. There was no effect of RNAi treatment on hemolymph glucose level or molt increment. Through multigene silencing and subsequent annotation of gene networks, gene blocks may provide a tailored approach to investigate complex polygenic traits with RNAi in a more efficient and scalable manner.
Details
- Title
- Silencing Multiple Crustacean Hyperglycaemic Hormone-Encoding Genes in the Redclaw Crayfish Cherax quadricarinatus Induces Faster Molt Rates with Anomalies
- Authors
- Nickolis Black - University of the Sunshine Coast, Queensland, Centre for BioinnovationThomas M. Banks - University of the Sunshine Coast, Queensland, Centre for BioinnovationSusan Glendinning - University of the Sunshine Coast, Queensland, Centre for BioinnovationGourab Chowdhury - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringDonald L. Mykles - Colorado State UniversityTomer Ventura (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
- Publication details
- International Journal of Molecular Sciences, Vol.25(22), pp.1-18
- Publisher
- MDPI AG
- Date published
- 2024
- DOI
- 10.3390/ijms252212314
- ISSN
- 1422-0067; 1661-6596
- Copyright note
- © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
- Data Availability
- RNA-Seq data are available online at NCBI Sequence Read Archive with the following accession number: SAMN42566432. Additional data are contained within Supplementary Materials.
- Grant note
- This work was supported by the National Science Foundation (IOS-1922701).
- Organisation Unit
- School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 991081598702621
- Output Type
- Journal article
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