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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

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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.

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Acknowledgments

This work was partially funded by the USDA-Biotechnology Risk Assessment Program (Grant No: 2009-33522-05774) and the Alabama Agricultural Experiment Station. The authors would like to thank Dr. Ron Thresher at CSIRO Marine and Atmospheric Research, Hobart, Tasmania, Australia for the review and comments on the first draft.

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Author notes

  1. Dayan A. Perera

    Present address: Gus R. Douglass Land-Grant Institute, West Virginia State University, Institute, WV, 25112, USA

  2. Carl A. Pinkert

    Present address: Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL, 35487, USA

  3. Jawahar G. Patil

    Present address: Fisheries and Aquaculture Centre, Institute for Marine and Antarctic Studies, University of Tasmania, Locked Bag, 1370, Launceston, TAS, 7250, Australia

Authors and Affiliations

  1. School of Fisheries, Aquaculture, and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA

    Baofeng Su, Mei Shang, Chao Li, Dayan A. Perera, Eric Peatman & Rex A. Dunham

  2. Department of Pathobiology, Auburn University, Auburn, AL, 36849, USA

    Carl A. Pinkert & Michael H. Irwin

  3. CSIRO, Marine and Atmospheric Research, Hobart, TAS, 7000, Australia

    Peter Grewe & Jawahar G. Patil

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  1. Baofeng Su
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Correspondence to Rex A. Dunham.

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Baofeng Su and Mei Shang are co-first authors.

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Su, B., Shang, M., Li, C. et al. Effects of transgenic sterilization constructs and their repressor compounds on hatch, developmental rate and early survival of electroporated channel catfish embryos and fry. Transgenic Res 24, 333–352 (2015). https://doi.org/10.1007/s11248-014-9846-4

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