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Effects of transgenic sterilization constructs and their repressor compounds on hatch, developmental rate and early survival of electroporated channel catfish embryos and fry

Citation

Su, B and Shang, M and Li, C and Perera, DA and Pinkert, CA and Irwin, MH and Peatman, E and Grewe, P and Patil, JG and Dunham, RA, Effects of transgenic sterilization constructs and their repressor compounds on hatch, developmental rate and early survival of electroporated channel catfish embryos and fry, Transgenic Research, 24, (2) pp. 333-352. ISSN 0962-8819 (2015) [Refereed Article]

Copyright Statement

Copyright 2014 Springer International

DOI: doi:10.1007/s11248-014-9846-4

Abstract

Channel catfish (Ictalurus punctatus) embryos were electroporated with sterilization constructs targeting primordial germ cell proteins or with buffer. Some embryos then were treated with repressor compounds, cadmium chloride, copper sulfate, sodium chloride or doxycycline, to prevent expression of the transgene constructs. Promoters included channel catfish nanos and vasa, salmon transferrin (TF), modified yeast Saccharomyces cerevisiae copper transport protein (MCTR) and zebrafish racemase (RM). Knock-down systems were the Tet-off (nanos and vasa constructs), MCTR, RM and TF systems. Knock-down genes included shRNAi targeting 5' nanos (N1), 3' nanos (N2) or dead end (DND), or double-stranded nanos RNA (dsRNA) for overexpression of nanos mRNA. These constructs previously were demonstrated to knock down nanos, vasa and dead end, with the repressors having variable success. Exogenous DNA affected percentage hatch (% hatch), as all 14 constructs, except for the TF dsRNA, TF N1 (T), RM DND (C), vasa DND (C), vasa N1 (C) and vasa N2 (C), had lower % hatch than the control electroporated with buffer. The MCTR and RM DND (T) constructs resulted in delayed hatch, and the vasa and nanos constructs had minimal effects on time of hatch (P<0.05). Cadmium chloride appeared to counteract the slow development caused by the TF constructs in two TF treatments (P\0.05). The 4 ppt sodium chloride treatment for the RM system decreased % hatch (P<0.05) and slowed development. In the case of nanos constructs, doxycycline greatly delayed hatch (P<0.05). Adverse effects of the transgenes and repressors continued for several treatments for the first 6 days after hatch, but only in a few treatments during the next 10 days. Repressors and gene expression impacted the yield of putative transgenic channel catfish fry, and need to be considered and accounted for in the hatchery phase of producing transgenically sterilized catfish fry and their fertile counterparts. This fry output should be considered to ensure that sufficient numbers of transgenic fish are produced for future applications and for defining repressor systems that are the most successful.

Item Details

Item Type:Refereed Article
Keywords:transgenics, catfish, sterile
Research Division:Agricultural and Veterinary Sciences
Research Group:Fisheries Sciences
Research Field:Aquaculture
Objective Division:Animal Production and Animal Primary Products
Objective Group:Fisheries - Aquaculture
Objective Field:Aquaculture Fin Fish (excl. Tuna)
Author:Patil, JG (Dr Jawahar Patil)
ID Code:96684
Year Published:2015
Web of Science® Times Cited:1
Deposited By:NC Marine Conservation and Resource Sustainability
Deposited On:2014-11-17
Last Modified:2017-11-04
Downloads:0

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