Dietary uptake of green fluorescent protein for delivery of dsRNA to induce RNA interference

La Fauce, Kathy, and Owens, Leigh (2012) Dietary uptake of green fluorescent protein for delivery of dsRNA to induce RNA interference. Journal of Marine Science Research and Development, 2 (4). 1000114. pp. 1-5.

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Abstract

Double stranded RNA (dsRNA) is a potent initiator of the RNA interference (RNAi) pathways in a diverse range of organisms including plants, invertebrates and vertebrates. RNAi is increasingly being investigated as an antiviral mechanism, but often the research is hampered by the safe and cost effective delivery of the dsRNA to the target cells. This study uses specifically designed E. coli-expressed plasmids containing green fluorescent protein (gfp), to determine if the plasmids still inside the bacterial cells can be successfully incorporated into host cells of the house cricket (Acheta domesticus), following ingestion of feed or water. This approach was trialled to determine, if using similar designed plasmids containing siRNAs can be effectively administered and provide protection against a target pathogen. Daily exposure through feed is the most effective for gfp expression (78%-91.8%) in A. domesticus, although weekly exposure was statistically equivalent and almost as good (75%-99.9%). Exposure via water gave reasonable uptake either daily or weekly (60%-84%, 14%-56%, respectively), but intensity was reduced and muscles did not demonstrate uptake (P<0.002). Continual or semi-continual exposure to gfp expressing bacteria resulted in increased uptake within cricket tissues, and increasing fluorescence (+++) with increasing exposure. Plasmid-based delivery via bacterial cells is therefore, an effective delivery tool and incorporation into feed allows a cost-effective method of providing continuous exposure for therapeutic purposes.

Item ID: 25394
Item Type: Article (Research - C1)
ISSN: 2155-9910
Keywords: RNA interference, bacterially expresses dsRNA, delivery, green fluorescent protein
Funders: Australia-India Strategic Research Fund
Date Deposited: 03 Apr 2013 06:36
FoR Codes: 06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060506 Virology @ 50%
07 AGRICULTURAL AND VETERINARY SCIENCES > 0704 Fisheries Sciences > 070404 Fish Pests and Diseases @ 50%
SEO Codes: 83 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 8301 Fisheries - Aquaculture > 830199 Fisheries - Aquaculture not elsewhere classified @ 100%
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