Abstract:Gene editing and gene silencing techniques have the potential to revolutionize our knowledge of biology and diseases of fish and other aquatic animals. By using such techniques, it is feasible to change the phenotype and modify cells, tissues and organs of animals in order to cure abnormalities and dysfunctions in the organisms. Gene editing is currently experimental in wide fields of aquaculture, including growth, controlled reproduction, sterility and disease resistance. Zink finger nucleases, TALENs and CRI… Show more
“…The evaluation of Interferons (IFNs) and IFN-stimulated genes by qPCR after Poly I:C stimulation showed an inhibition of the expression in the mavs and irf3 edited groups, showing the advantages of using CRISPR edited cell lines in fish for understanding the immune response and the host-pathogen interaction. While the CRISPR-Cas system has been successfully used in different fish species, including S. salar, Oreochromis niloticus, Danio rerio, and Oryzias latipes [29], most studies using the CRISPR-Cas system in fish have focused on evaluating the effects of the edition in the target genes and/or its directly related genes (i.e., receptors and transcription factors) [30,31], without considering the global impact on gene expression inside the cell, thereby missing information that could be relevant in the interpretation of experimental results. As observed in our results, RNA-seq data showed relevant significant differences in general transcriptional regulation between TIII and TVIII targets, even when no relevant differences were observed in the expression of transferrin and transferrin-related genes.…”
Section: Fish Cell Lines and Crispr-cas9 System As Genome-editing Too...mentioning
In this study, we investigated the function of a gene associated with iron metabolism using CRISPR-Cas9 and RNA sequencing in SHK-1 salmon cells. Our objective was to understand how different guide RNA (gRNA) sequences against the transferrin gene tf could influence gene expression and cellular processes related to iron uptake. RNA-Seq analysis was performed to evaluate the transcriptomic effects of two distinct gRNA targets with high knock-out (KO) efficiencies for the targeted tf gene in the SHK-1 genome. Our results showed no significant differential expression in transferrin-related transcripts between wild-type and CRISPR-edited cells; however, there were major differences between their transcriptomes, indicating complex transcriptional regulation changes. Enrichment analysis highlighted specific processes and molecular functions, including those related to the nucleus, cytoplasm, and protein binding. Notably, different sgRNAs targeting tf might result in different mutations at DNA levels in SHK-1 salmon cells.
“…The evaluation of Interferons (IFNs) and IFN-stimulated genes by qPCR after Poly I:C stimulation showed an inhibition of the expression in the mavs and irf3 edited groups, showing the advantages of using CRISPR edited cell lines in fish for understanding the immune response and the host-pathogen interaction. While the CRISPR-Cas system has been successfully used in different fish species, including S. salar, Oreochromis niloticus, Danio rerio, and Oryzias latipes [29], most studies using the CRISPR-Cas system in fish have focused on evaluating the effects of the edition in the target genes and/or its directly related genes (i.e., receptors and transcription factors) [30,31], without considering the global impact on gene expression inside the cell, thereby missing information that could be relevant in the interpretation of experimental results. As observed in our results, RNA-seq data showed relevant significant differences in general transcriptional regulation between TIII and TVIII targets, even when no relevant differences were observed in the expression of transferrin and transferrin-related genes.…”
Section: Fish Cell Lines and Crispr-cas9 System As Genome-editing Too...mentioning
In this study, we investigated the function of a gene associated with iron metabolism using CRISPR-Cas9 and RNA sequencing in SHK-1 salmon cells. Our objective was to understand how different guide RNA (gRNA) sequences against the transferrin gene tf could influence gene expression and cellular processes related to iron uptake. RNA-Seq analysis was performed to evaluate the transcriptomic effects of two distinct gRNA targets with high knock-out (KO) efficiencies for the targeted tf gene in the SHK-1 genome. Our results showed no significant differential expression in transferrin-related transcripts between wild-type and CRISPR-edited cells; however, there were major differences between their transcriptomes, indicating complex transcriptional regulation changes. Enrichment analysis highlighted specific processes and molecular functions, including those related to the nucleus, cytoplasm, and protein binding. Notably, different sgRNAs targeting tf might result in different mutations at DNA levels in SHK-1 salmon cells.
“…Gene editing, particularly through the CRISPR/Cas9 system, has revolutionized the precision with which we can alter the genetic makeup of organisms. This technology allows for the targeted modification of DNA to include point mutations, gene knockouts, and large-scale genomic rearrangements, thereby enabling researchers to directly assess gene function and trait manifestation [153][154][155].…”
Fish exhibit a broad spectrum of colors and patterns facilitated by specialized cells known as chromatophores. The vibrant coloration of fish, controlled by complex genetic and environmental interactions, serves critical roles in ecological functions such as mating, predation, and camouflage. This diversity not only makes fish an invaluable model for exploring the molecular mechanisms of pigmentation but also significantly impacts their economic value within the aquaculture industry, where color traits can drive marketability and breeding choices. This review delves into the sophisticated biological processes governing fish pigmentation and discusses their applications in enhancing aquaculture practices. By exploring the intersection of genetic regulation, environmental influences, and advanced breeding techniques, this review highlights both the scientific understanding and practical applications of fish coloration, providing a bridge between basic biological research and its application in commercial aquaculture.
“…Applied to tilapia, genome editing can develop solutions for aquaculture challenges and investigate the genetic, developmental, and physiological mechanisms underlying commercially relevant traits [ 123 ]. Genome editing for aquaculture purposes has been widely reviewed [ 72 , 124 – 126 ] including from biosafety, sustainability, and welfare perspectives [ 127 – 129 ] so here we focus on studying the development and evolution of phenotypes. Figure 3 Overview of the state-of-the-art and the emerging future of genome editing in cichlids.…”
Section: From the State-of-the-art To New Frontiers In Cichlid Genome...mentioning
African cichlid fishes of the Cichlidae family are a group of teleosts important for aquaculture and research. A thriving research community is particularly interested in the cichlid radiations of the East African Great Lakes. One key goal is to pinpoint genetic variation underlying phenotypic diversification, but the lack of genetic tools has precluded thorough dissection of the genetic basis of relevant traits in cichlids. Genome editing technologies are well established in teleost models like zebrafish and medaka. However, this is not the case for emerging model organisms, such as East African cichlids, where these technologies remain inaccessible to most laboratories, due in part to limited exchange of knowledge and expertise. The Cichlid Science 2022 meeting (Cambridge, UK) hosted for the first time a Genome Editing Workshop, where the community discussed recent advances in genome editing, with an emphasis on CRISPR/Cas9 technologies. Based on the workshop findings and discussions, in this review we define the state-of-the-art of cichlid genome editing, share resources and protocols, and propose new possible avenues to further expand the cichlid genome editing toolkit.
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