Mar. Drugs 2017, 15(6), 155; https://doi.org/10.3390/md15060155
Salt Sensitive Tet-Off-Like Systems to Knockdown Primordial Germ Cell Genes for Repressible Transgenic Sterilization in Channel Catfish, Ictalurus punctatus
Hanbo Li 1,†, Baofeng Su 1,2,†, Guyu Qin 1
, Zhi Ye 1, Ahmed Alsaqufi 1,3, Dayan A. Perera 1,4, Mei Shang 1,2, Ramjie Odin 1, Khoi Vo 1, David Drescher 1, Dalton Robinson 1, Dan Zhang 1, Nermeen Abass 1,5 and Rex A. Dunham 1,*
, Zhi Ye 1, Ahmed Alsaqufi 1,3, Dayan A. Perera 1,4, Mei Shang 1,2, Ramjie Odin 1, Khoi Vo 1, David Drescher 1, Dalton Robinson 1, Dan Zhang 1, Nermeen Abass 1,5 and Rex A. Dunham 1,*
1
School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL 36849, USA
2
National and Local United Engineering Laboratory for Freshwater Fish Breeding, Heilongjiang River Fisheries Research Institute, Chinese Academy of Fisheries Sciences, Harbin 150070, China
3
Department of Aquaculture and Animal production, King Faisal University, Al Ahsa 31982, Saudi Arabia
4
Research and Development Corporation, Gus R. Douglass Institute, West Virginia State University, WV 25112, USA
5
Department of Agricultural Botany, Faculty of Agriculture Saba-Basha, Alexandria University, Alexandria City, P.O. Box 2153, Egypt
†
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editors: Allen Place, Rosemary Jagus, Joy Watts and Paul Long
Received: 1 April 2017 / Revised: 22 May 2017 / Accepted: 25 May 2017 / Published: 31 May 2017
(This article belongs to the Special Issue Advances and New Perspectives in Marine Biotechnology II 2016)
Abstract
Repressible knockdown approaches were investigated for transgenic sterilization in channel catfish, Ictalurus punctatus. Two primordial germ cell (PGC) marker genes, nanos and dead end, were targeted for knockdown, and an off-target gene, vasa, was monitored. Two potentially salt sensitive repressible promoters, zebrafish adenylosuccinate synthase 2 (ADSS) and zebrafish racemase (Rm), were each coupled with four knockdown strategies: ds-sh RNA targeting the 5′ end (N1) or 3′ end (N2) of channel catfish nanos, full-length cDNA sequence of channel catfish nanos for overexpression (cDNA) and ds-sh RNA targeting channel catfish dead end (DND). Each construct had an untreated group and treated group with sodium chloride as the repressor compound. Spawning rates of full-sibling P1 fish exposed or not exposed to the constructs as treated and untreated embryos were 93% and 59%, respectively, indicating potential sterilization of fish and repression of the constructs. Although the mRNA expression data of PGC marker genes were inconsistent in P1 fish, most F1 individuals were able to downregulate the target genes in untreated groups and repress the knockdown process in treated groups. The results indicate that repressible transgenic sterilization is feasible for reproductive control of fish, but more data from F2 or F3 are needed for evaluation. View Full-Text
Figure 1
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Li, H.; Su, B.; Qin, G.; Ye, Z.; Alsaqufi, A.; Perera, D.A.; Shang, M.; Odin, R.; Vo, K.; Drescher, D.; Robinson, D.; Zhang, D.; Abass, N.; Dunham, R.A. Salt Sensitive Tet-Off-Like Systems to Knockdown Primordial Germ Cell Genes for Repressible Transgenic Sterilization in Channel Catfish, Ictalurus punctatus. Mar. Drugs 2017, 15, 155.
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