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141077 - Double-stranded RNA binding proteins in serum contribute to systemic RNAi across phyla.pdf (3.17 MB)

Double-stranded RNA binding proteins in serum contribute to systemic RNAi across phyla - towards finding the missing link in Achelata

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journal contribution
posted on 2023-05-20, 18:08 authored by Banks, TM, Wang, T, Quinn FitzgibbonQuinn Fitzgibbon, Gregory SmithGregory Smith, Ventura, T
RNA interference (RNAi) has become a widely utilized method for studying gene function, yet despite this many of the mechanisms surrounding RNAi remain elusive. The core RNAi machinery is relatively well understood, however many of the systemic mechanisms, particularly double-stranded RNA (dsRNA) transport, are not. Here, we demonstrate that dsRNA binding proteins in the serum contribute to systemic RNAi and may be the limiting factor in RNAi capacity for species such as spiny lobsters, where gene silencing is not functional. Incubating sera from a variety of species across phyla with dsRNA led to a gel mobility shift in species in which systemic RNAi has been observed, with this response being absent in species in which systemic RNAi has never been observed. Proteomic analysis suggested lipoproteins may be responsible for this phenomenon and may transport dsRNA to spread the RNAi signal systemically. Following this, we identified the same gel shift in the slipper lobster Thenus australiensis and subsequently silenced the insulin androgenic gland hormone, marking the first time RNAi has been performed in any lobster species. These results pave the way for inducing RNAi in spiny lobsters and for a better understanding of the mechanisms of systemic RNAi in Crustacea, as well as across phyla.

Funding

Australian Research Council

Orna-Tas Pty Ltd

University of the Sunshine Coast

History

Publication title

International Journal of Molecular Sciences

Volume

21

Issue

18

Article number

6967

Number

6967

Pagination

1-20

ISSN

1422-0067

Department/School

Institute for Marine and Antarctic Studies

Publisher

Molecular Diversity Preservation International

Place of publication

Matthaeusstrasse 11, Basel, Switzerland, Ch-4057

Rights statement

© 2020 by the authors. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) http://creativecommons.org/licenses/by/4.0/

Repository Status

  • Open

Socio-economic Objectives

Aquaculture crustaceans (excl. rock lobster and prawns); Aquaculture rock lobster

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