Double Transgenesis of Humanized fat1 and fat2 Genes Promotes Omega-3 Polyunsaturated Fatty Acids Synthesis in a Zebrafish Model

Pang, Shaochen; Wang, Hou-Peng; Li, Kuo-Yu; Zhang, Zhu; Kang, Jing; Sun, Yuhan

doi:10.1007/s10126-014-9577-9

Cited by 40 publications

(30 citation statements)

References 52 publications

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“…Transgenic zebrafish (Danio rerio) have been produced by transformation with Δ6 and Δ5 fatty acyl desaturases of masu salmon (Oncorhynchus masou) (Alimuddin et al, 2005(Alimuddin et al, , 2007, and both zebrafish (Alimuddin et al, 2008) and the marine teleost, nibe croaker Nibea mitsukurii (Kabeya et al, 2014), have been transformed with fatty acid elongase from masu salmon. Recently, the production of n-3 PUFA, specifically LNA from 18:1n-9, was demonstrated in GM-zebrafish transformed with fat-1 and fat-2 genes (Pang et al, 2014). However, in all these studies the increments in EPA and DHA levels have been small and so this enhancement, by a combination of adding function and/or overexpression of biosynthetic genes, may have limitations based on the metabolic and physiological controls of the biochemical pathways.…”

Section: Enhancing Endogenous Lc-pufa Production and Retentionmentioning

confidence: 99%

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

2015

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Abbreviations: ARA, arachidonic acid (20:4n-6); CVD, cardiovascular disease; DHA, docosahexaenoic acid (22:6n-3); DPA, docosapentaenoic acid (22:5n-3); EFA, essential fatty acid; Elovl, fatty acid elongase; EPA, eicosapentaenoic acid (20:5n-3); Fads, fatty acyl desaturase; FM, fishmeal; FO, fish oil; LC-PUFA, long-chain polyunsaturated fatty acids (≥ C20 ≥ 3 double bonds); LOA, linoleic acid (18:2n-6); LNA, linolenic acid (18:3n-3); PUFA, polyunsaturated fatty acid; TAG, triacylglycerol; VO, vegetable oil. ABSTRACTIn the 40 years since the essentiality of polyunsaturated fatty acids (PUFA) in fish was first established by determining quantitative requirements for 18:3n-3 and 18:2n-6 in rainbow trout, essential fatty acid (EFA) research has gone through distinct phases. For 20 years the focus was primarily on determining qualitative and quantitative EFA requirements of fish species. Nutritional and biochemical studies showed major differences between fish species based on whether C 18 PUFA or long-chain (LC)-PUFA were required to satisfy requirements. In contrast, in the last 20 years, research emphasis shifted to determining "optimal" levels of EFA to support growth of fish fed diets with increased lipid content and where growth expectations were much higher. This required greater knowledge of the roles and functions of EFA in metabolism and physiology, and how these impacted on fish health and disease. Requirement studies were more focused on early life stages, in particular larval marine fish, defining not only levels, but also balances between different EFA. Finally, a major driver in the last 10-15 years has been the unavoidable replacement of fish oil and fishmeal in feeds and the impacts that this can have on n-3 LC-PUFA contents of diets and farmed fish, and the human consumer. Thus, dietary n-3 in fish feeds can be defined by three levels.Firstly, the minimum level required to satisfy EFA requirements and thus prevent nutritional pathologies. This level is relatively small and easy to supply even with today's current high demand for fish oil. The second level is that required to sustain maximum growth and optimum health in fish being fed modern high-energy diets. The balance between different PUFA and LC-PUFA is important and defining them is more challenging, and so ideal levels and balances are still not well understood, particularly in relation to fish health. The third level is currently driving much research; how can we supply sufficient n-3 LC-PUFA to maintain the nutritional quality of farmed fish at the same level as 20 years ago, and similar or better than in wild fish? This level far exceeds the biological requirements of the fish itself and to satisfy it we require entirely new sources of n-3 LC-PUFA. We cannot rely on the finite and limited marine resources that we can sustainably harvest or 3 efficiently recycle. We need to produce n-3 LC-PUFA de novo and all possible options should be considered.Keywords: Fish oil; essential fatty acids; nutrition; health; metabolism; sustainability 4 Highl...

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Section: Enhancing Endogenous Lc-pufa Production and Retentionmentioning

confidence: 99%

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

2015

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“…Thus, feeding costs are increased because of the need to supplement diets with extra n3 or n6 PUFA. Recently, de novo LC-PUFA biosynthesis was induced in zebrafish by using fat1 and fat1/fat2 double transgenic fish, resulting in robust n-3 LC-PUFA production even when using low PUFA food [67]. Transgenic fish can also be used as a bioreactor.…”

Section: Transgenic Breedingmentioning

confidence: 99%

Fish genome manipulation and directional breeding

Zhu

Sun

2015

Sci. China Life Sci.

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Aquaculture is one of the fastest developing agricultural industries worldwide. One of the most important factors for sustainable aquaculture is the development of high performing culture strains. Genome manipulation offers a powerful method to achieve rapid and directional breeding in fish. We review the history of fish breeding methods based on classical genome manipulation, including polyploidy breeding and nuclear transfer. Then, we discuss the advances and applications of fish directional breeding based on transgenic technology and recently developed genome editing technologies. These methods offer increased efficiency, precision and predictability in genetic improvement over traditional methods. fish directional breeding, polyploidy breeding, nuclear transfer, transgenic fish, genome editing Citation:Ye D, Zhu ZY, Sun YH. Fish genome manipulation and directional breeding. Sci China Life Sci, 2015Sci, , 58: 170 -177, doi: 10.1007 Fish are one of the most important sources of protein for humans. As a result of declines in wild fisheries, aquaculture has become one of the fastest developing agricultural industries worldwide [1]. To increase the sustainability of aquaculture, culturists commonly conduct selective breeding to develop strains that perform well in captivity. Unfortunately, the sustainability of many sectors of the industry is negatively affected by inbreeding depression, disease outbreaks, under production, and low meat quality [2]. To address these issues, there is an urgent need for the development of high quality fish strains that have high growth rates, disease-resistance, and/or higher nutritional value.Traditional crossbreeding methods such as intra-species crossbreeding [3] and inter-species hybridization [4] have been successfully used for several decades. However, crossbreeding requires multiple-generations of hybridization to introduce a desirable trait to a given strain. Additionally, the outcomes of these methods are unpredictable because the underlying mechanisms controlling desirable traits are unknown. Thus, there is a need to develop more efficient, precise and predictable techniques for producing production scale numbers of high-quality fish. We review the history of fish breeding methods based on classical genome manipulation approaches, including polyploidy breeding [5] and nuclear transfer [6]. Then, we discuss directional breeding of fish based on transgenic technology and recently developed genome editing technologies. Advances in breeding methodology based on classical genome manipulation and recently developed genome editing methods will likely play a major role in the future of genetic breeding in fish.

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“…The EGFP-expressing embryos were raised, and transgenic lines were generated as described previously (Pang et al 2014).…”

Section: Generation Of Transgenic Zebrafishmentioning

confidence: 99%

“…The cDNAs of embryos, larvae, and muscle tissue were synthesized as described (Pang et al 2014). Quantitative RT-PCR was performed using SYBR Mix (Bio-Rad) and a CFX RealTime PCR System (Bio-Rad).…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

Transcriptional Activity and DNA Methylation Dynamics of the Gal4/UAS System in Zebrafish

et al. 2015

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The Gal4/upstream activating sequence (UAS) system is a powerful genetic tool for the temporal and spatial expression of target genes. In this study, the dynamic activity of the Gal4/UAS system was monitored in zebrafish throughout the entire lifespan and during germline transmission, using an optimized Gal4/UAS, KalTA4/4xUAS, which is driven by two muscle-specific regulatory sequences. We found that UAS-linked gene expression was transcriptionally amplified by Gal4/UAS during early developmental stages and that the amplification effects tended to weaken during later stages and even disappear in subsequent generations. In the F2 generation, the transcription of a UAS-linked enhanced green fluorescent protein (EGFP) reporter was transcriptionally silent from 16 days post-fertilization (dpf) into adulthood, yet offspring of this generation showed reactivation of the EGFP reporter in some strains. We further show that the transcriptional silencing and reactivation of UAS-driven EGFP correlated with the DNA methylation levels of the UAS regulatory sequences. Notably, asymmetric DNA methylation of the 4xUAS occurred in oocytes and sperm. Moreover, the paternal and maternal 4xUAS sequences underwent different DNA methylation dynamics after fertilization. Our study suggests that the Gal4/UAS system may represent a powerful tool for tracing the DNA methylation dynamics of paternal and maternal loci during zebrafish development and that UASspecific DNA methylation should be seriously considered when the Gal4/UAS system is applied in zebrafish.

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Double Transgenesis of Humanized fat1 and fat2 Genes Promotes Omega-3 Polyunsaturated Fatty Acids Synthesis in a Zebrafish Model

Cited by 40 publications

References 52 publications

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective

Fish genome manipulation and directional breeding

Transcriptional Activity and DNA Methylation Dynamics of the Gal4/UAS System in Zebrafish

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