CRISPR/Cas9-mediated precision integration of fat-1 and fat-2 from Caenorhabditis elegans at long repeated sequence in channel catfish (Ictalurus punctatus) and the impact on n-3 fatty acid level

Xing, De; Shang, Mei; Li, Shangjia; Wang, Wenwen; Hasin, Tasnuba; Hettiarachchi, Darshika; Alston, Veronica; Bern, Logan; Qin, Zhenkui; Su, Baofeng; Dunham, Rex A.

doi:10.1016/j.aquaculture.2023.739229

Cited by 7 publications

(3 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A behavioural trait targeted in Pacific bluefin tuna could be identified: GM tuna showed slower swimming behaviour, which might reduce losses of GM tuna from sea cages to the open sea ( Higuchi et al, 2019 ). In order to improve product quality, especially meat quality, the content of n-3 PUFAs is increased in channel catfish ( Xing et al, 2022 ; Xing et al, 2023 ). The same trait is altered in terrestrial farm animals (see previous section) and in the model fish D. rerio ( Pang et al, 2014 ).…”

Section: Resultsmentioning

confidence: 99%

Horizon scanning of potential environmental applications of terrestrial animals, fish, algae and microorganisms produced by genetic modification, including the use of new genomic techniques

Miklau,

Burn,

Eckerstorfer

et al. 2024

Front. Genome Ed.

View full text Add to dashboard Cite

With scientific progress and the development of new genomic techniques (NGTs), the spectrum of organisms modified for various purposes is rapidly expanding and includes a wide range of taxonomic groups. An improved understanding of which newly developed products may be introduced into the market and released into the environment in the near and more distant future is of particular interest for policymakers, regulatory authorities, and risk assessors. To address this information need, we conducted a horizon scanning (HS) of potential environmental applications in four groups of organisms: terrestrial animals (excluding insects and applications with gene drives), fish, algae and microorganisms. We applied a formal scoping review methodology comprising a structured search of the scientific literature followed by eligibility screening, complemented by a survey of grey literature, and regulatory websites and databases. In all four groups of organisms we identified a broad range of potential applications in stages of basic as well as advanced research, and a limited number of applications which are on, or ready to be placed on, the market. Research on GM animals including fish is focused on farmed animals and primarily targets traits which increase performance, influence reproduction, or convey resistance against diseases. GM algae identified in the HS were all unicellular, with more than half of the articles concerning biofuel production. GM algae applications for use in the environment include biocontrol and bioremediation, which are also the main applications identified for GM microorganisms. From a risk assessor’s perspective these potential applications entail a multitude of possible pathways to harm. The current limited level of experience and limited amount of available scientific information could constitute a significant challenge in the near future, for which risk assessors and competent authorities urgently need to prepare.

show abstract

Section: Resultsmentioning

confidence: 99%

Horizon scanning of potential environmental applications of terrestrial animals, fish, algae and microorganisms produced by genetic modification, including the use of new genomic techniques

Miklau,

Burn,

Eckerstorfer

et al. 2024

Front. Genome Ed.

View full text Add to dashboard Cite

show abstract

“…Beyond the successful achievement of one desired trait, it is possible to use CRISPR/Cas9 to simultaneously improve multiple characteristics based on empirical data and theoretical foundations, 89–92 which means that we could alter other traits through the construction of different vectors while we focus on enhancement of disease resistance. Publications have already shown that single or multiple transgenes can be integrated into the fish genome by single sgRNA‐mediated genome editing (ssGE) or multi‐sgRNA‐mediated multiple genome editing assisted using plasmid donors 92,93 . Wang et al 34 demonstrated that the cathelicidin transgene can be knocked into the LH locus of channel catfish by double‐stranded DNA‐mediated ssGE.…”

Section: Future Perspectivesmentioning

confidence: 99%

“…Publications have already shown that single or multiple transgenes can be integrated into the fish genome by single sgRNA-mediated genome editing (ssGE) or multi-sgRNA-mediated multiple genome editing assisted using plasmid donors. 92,93 Wang et al 34 demonstrated that the cathelicidin transgene can be knocked into the LH locus of channel catfish by double-stranded DNA-mediated ssGE. In addition, a double-cut plasmid containing a foreign AMG transgene as a donor is beneficial to improve integration rates using CRISPR/Cas9-mediated ssGE.…”

Section: Transgenic Amgs and Fish Immune Systemmentioning

confidence: 99%

Enhancing aquaculture disease resistance: Antimicrobial peptides and gene editing

Wang

Cheng

2023

Reviews in Aquaculture

View full text Add to dashboard Cite

Previous studies have demonstrated that CRISPR/Cas9‐mediated genome editing and transgenesis by integrating vector‐engineered antimicrobial peptide genes (AMGs) are effective to modulate the fish's innate immune system. To generalise the knowledge of AMG application in aquaculture, we recruited 544 data entries from a pool of empirical studies, which included 23 studies (two unpublished from our team) spanning 12 diseases. We systematically re‐processed and re‐analysed these data by harnessing a cross‐disease meta‐analysis. On aggregate, AMG‐genetic engineering aimed at enhancing disease resistance was shown to decrease the number of colony‐forming units of bacteria from fish tissues, increase post‐infection survival rates, and alter the expression of AMGs and immune‐related genes. Further, AMG pathogen‐combating activity was triggered within 2 h after infection and lasted 48 h. The overexpression of AMGs was highest in the spleen and skin, followed by the kidney and liver during this period. Typically, regardless of the type of AMGs, the synergistic expression of AMGs with IL‐10, IKβ, TGFβ, C3b and TLR genes in AMG‐integrated fish contributed to activating inflammatory/immune responses against pathogens. Our findings revealed that the efficiency of transgenic AMGs against pathogens was fish‐, pathogen‐ and AMG‐specific. Compared to bacteria, transgenic AMGs were less inhibitory to viruses and parasites. In addition, innovative CRISPR/Cas9‐based transgenesis enabling the site‐directed knock‐in of foreign genes at multiple loci was assessed for the enhancement of disease resistance in combination with other favourable fish‐production traits, including fast growth, sterility and enriched fatty‐acid content. Altogether, our findings indicated that AMGs as transgenes have substantial promise to modulate the fish's innate immune system and enhance disease resistance.

show abstract

Chitosan-based delivery of fish codon-optimised Caenorhabditis elegans FAT-1 and FAT-2 boosts EPA and DHA biosynthesis in Sparus aurata

Wu,

Rashidpour,

Fàbregas

et al. 2024

Rev Fish Biol Fisheries

View full text Add to dashboard Cite

Omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) are essential fatty acids required in healthy balanced diets for humans. To induce sustained production of n-3 LC-PUFA in gilthead seabream (Sparus aurata), chitosan-tripolyphosphate (TPP) nanoparticles encapsulating plasmids expressing fish codon-optimised Caenorhabditis elegans FAT-1 and FAT-2 were intraperitoneally administered every 4 weeks (3 doses in total, each of 10 μg plasmid per g of body weight). Growth performance and metabolic effects of chitosan-TPP complexed with pSG5 (empty plasmid), pSG5-FAT-1, pSG5-FAT-2 and pSG5-FAT-1 + pSG5-FAT-2 were assessed 70 days post-treatment. Tissue distribution analysis showed high expression levels of fish codon-optimised FAT-1 and FAT-2 in the liver (> 200-fold). Expression of fat-1 and fat-1 + fat-2 increased weight gain. Fatty acid methyl esters assay revealed that co-expression of fat-1 and fat-2 increased liver production and muscle accumulation of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and total n-3 LC-PUFA, while decreased the n-6/n-3 ratio. Co-expression of fat-1 and fat-2 downregulated srebf1 and genes encoding rate-limiting enzymes for de novo lipogenesis in the liver, leading to decreased circulating triglycerides and cholesterol. In contrast, FAT-2 and FAT-1 + FAT-2 upregulated hepatic hnf4a, nr1h3 and key enzymes in glycolysis and the pentose phosphate pathway. Our findings demonstrate for the first time efficient and sustained production of EPA and DHA in animals after long-term treatment with chitosan-TPP-DNA nanoparticles expressing FAT-1 and FAT-2, which enabled the production of functional fish rich in n-3 LC-PUFA for human consumption.

show abstract

CRISPR/Cas9-mediated precision integration of fat-1 and fat-2 from Caenorhabditis elegans at long repeated sequence in channel catfish (Ictalurus punctatus) and the impact on n-3 fatty acid level

Cited by 7 publications

References 47 publications

Horizon scanning of potential environmental applications of terrestrial animals, fish, algae and microorganisms produced by genetic modification, including the use of new genomic techniques

Horizon scanning of potential environmental applications of terrestrial animals, fish, algae and microorganisms produced by genetic modification, including the use of new genomic techniques

Enhancing aquaculture disease resistance: Antimicrobial peptides and gene editing

Chitosan-based delivery of fish codon-optimised Caenorhabditis elegans FAT-1 and FAT-2 boosts EPA and DHA biosynthesis in Sparus aurata

Contact Info

Product

Resources

About