Crustacean Nuclear Localization Signals Help Facilitating the Delivery of DNA into Australian Red-claw Crayfish Cells

Chan D H Nguyen; Tomer Ventura; Abigail Elizur

doi:10.1016/j.aquaculture.2018.09.030

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Crustacean Nuclear Localization Signals Help Facilitating the Delivery of DNA into Australian Red-claw Crayfish Cells

Journal article

Open access

Peer reviewed

Crustacean Nuclear Localization Signals Help Facilitating the Delivery of DNA into Australian Red-claw Crayfish Cells

Chan D H Nguyen, Tomer Ventura and Abigail Elizur

Aquaculture, Vol.499, pp.149-159

2019

DOI: https://doi.org/10.1016/j.aquaculture.2018.09.030

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Abstract

DNA delivery

nuclear localisation signal

NLS

Australian redclaw crayfish

Cherax quadricarinatus

Crustacean aquaculture is rapidly expanding, and demands are supported through the application of novel technologies. One such application is the oral administration of foreign DNA, which expresses genes of interest to promote growth, health and superior quality traits. To date, there has not been any report of a viable method for large-scaled oral delivery of foreign DNA into crustaceans, although these methodologies have expanded in utilization across many plant and animal species. DNA delivery requires a thorough understanding of the transcription machinery of the host species to which it is administered. This knowledge is still unavailable in crustaceans, where the mechanism to enable nuclear entry is obscure. In this study we utilized a bioinformatics approach to shortlist Nuclear Localisation Signals (NLSs), and designed crustacean-specific NLSs to enhance the delivery of DNA into the nucleus via active transport. The NLS-plasmid complex, carrying GFP and IAG genes was transfected into Australian redclaw crayfish (Cherax quadricarinatus) primary cultured cells, resulting in protein expression of the introduced genes. These NLSs also increased the transfection efficiency in mammalian cells from 0.6% to 9% in suboptimal conditions. Hence, this integrated approach of bioinformatics and in vitro studies could help in developing a novel biotechnology to enable the large-scaled oral DNA delivery in the crustacean aquaculture industry.

Details

Title: Crustacean Nuclear Localization Signals Help Facilitating the Delivery of DNA into Australian Red-claw Crayfish Cells
Authors: Chan D H Nguyen (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Tomer Ventura (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Abigail Elizur (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Publication details: Aquaculture, Vol.499, pp.149-159
Publisher: Elsevier BV
Date published: 2019
DOI: 10.1016/j.aquaculture.2018.09.030
ISSN: 0044-8486
Grants: Redefining the molecular mechanism underlying crustacean metamorphosis, DP160103320, Australian Research Council (Australia, Canberra) - ARC
Organisation Unit: School of Science and Engineering - Legacy; University of the Sunshine Coast, Queensland; GeneCology Research Centre - Legacy; School of Science, Technology and Engineering; Centre for Bioinnovation
Language: English
Record Identifier: 99450879702621
Output Type: Journal article

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