Genomics and Applied Biology 2024, Vol.15, No.2, 89-98 http://bioscipublisher.com/index.php/gab 89 Research Insight Open Access Efficient Gene Transfer Techniques in Shrimp and Their Cellular Applications Fei Zhao, Fan Wang Aquatic Biology Research Center, Cuixi Academy of Biotechnology, Zhuji, 311800, Zhejiang, China Co-corresponding author: fan.wang@cuixi.org Genomics and Applied Biology, 2024, Vol.15, No.2 doi: 10.5376/gab.2024.15.0011 Received: 17 Feb., 2024 Accepted: 21 Mar., 2024 Published: 03 Apr., 2024 Copyright © 2024 Zhao and Wang, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Zhao F., and Wang F., 2024, Efficient gene transfer techniques in shrimp and their cellular applications, Genomics and Applied Biology, 15(2): 89-98 (doi: 10.5376/gab.2024.15.0011) Abstract This study compared the efficiency of microinjection, electroporation, and transfection methods for gene transfer into shrimp zygotes. Transfection using the jetPEI reagent demonstrated the highest efficiency, with hatching rates of 50%-60% and gene expression rates of 40%-60%. Additionally, a VP28-pseudotyped baculovirus system achieved up to 100% infection efficiency in adult shrimp tissues, although it exhibited tissue-specific tropism. A triple-pseudotyped retroviral system also showed promise, particularly in shrimp primary lymphoid cells, with infection efficiencies of 20%-30%. Furthermore, the inclusion of shrimp-specific promoters and viral envelope proteins significantly enhanced the tropism and infectivity of lentiviral vectors in shrimp cells. The findings indicate that transfection with jetPEI and the use of pseudotyped viral systems are highly effective for gene transfer in shrimp. These methods hold significant potential for advancing genetic manipulation and cellular studies in shrimp, which could lead to improved disease resistance and other desirable traits in aquaculture. Keywords Gene transfer; Shrimp; Transfection; Electroporation; Microinjection; Pseudotyped viral systems; jetPEI; VP28; Lentiviral vectors; Aquaculture 1 Introduction Gene transfer techniques have revolutionized the field of aquaculture, providing powerful tools for genetic modification and enhancement of various aquatic species. These techniques, including CRISPR/Cas9, TALENs, and Zink finger nucleases, have been widely applied to fish species to improve traits such as disease resistance, growth, and reproduction (Gutási et al., 2023). The application of these methods in shrimp, however, is still in its nascent stages, with significant potential for development. The ability to modify the genome of shrimp can lead to substantial improvements in aquaculture productivity and sustainability (Zenger et al., 2019; Jacinda and Yustiati, 2021). Shrimp aquaculture is a critical component of the global seafood industry, contributing significantly to food security and economic development. However, the industry faces challenges such as disease outbreaks and environmental sustainability. Gene transfer techniques offer a promising solution to these issues by enabling the development of disease-resistant shrimp strains and enhancing growth rates (Sun et al., 2005; Jacinda and Yustiati, 2021). For instance, the use of transfection methods has shown high efficiency in gene transfer, with significant improvements in hatching rates and gene expression in shrimp (Sun et al., 2005). Additionally, the identification of horizontally transferred genes in shrimp genomes highlights the potential for natural genetic enhancements that can be harnessed through gene transfer technologies (Yuan et al., 2013). This study aims to provide a comprehensive overview of the current state of gene transfer techniques in shrimp, their applications, and potential benefits. The objectives are to review the various gene transfer methods used in shrimp, including microinjection, electroporation, and transfection, discuss the efficiency and outcomes of these methods in terms of gene expression and phenotypic changes, explore the potential applications of gene transfer in improving shrimp aquaculture, focusing on disease resistance, growth enhancement, and environmental sustainability, and address the ethical and environmental considerations associated with the use of genetically modified organisms (GMOs) in aquaculture.
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