Genomics and Applied Biology 2024, Vol.15, No.2, 89-98 http://bioscipublisher.com/index.php/gab 91 3.1.3 Lipofection Lipofection uses lipid-based reagents to form complexes with nucleic acids, facilitating their entry into cells. This method is less commonly used in shrimp but has potential due to its non-invasive nature and ease of use. Lipofection is particularly useful for in vitro applications and can be optimized for different cell types and conditions. 3.1.4 Viral Vectors Viral vectors are engineered viruses that can deliver genetic material into host cells. This method is highly efficient and can achieve stable gene expression. Viral vectors have been used in various model organisms and hold promise for shrimp gene transfer due to their ability to infect a wide range of cell types and integrate genetic material into the host genome. 3.2 Comparative analysis of techniques Each gene transfer technique has its advantages and limitations. Microinjection is highly precise but labor-intensive and requires specialized equipment (Abdelrahman et al., 2021; Lane et al., 2021; Gultom, 2023). Electroporation is less invasive and suitable for large-scale applications but may have lower efficiency in some cases (Takahashi et al., 2015). Lipofection is easy to use and non-invasive but may not be as effective in vivo. Viral vectors offer high efficiency and stable gene expression but pose biosafety concerns and require careful handling. 3.3 Efficiency and success rates in shrimp The efficiency and success rates of gene transfer techniques in shrimp vary depending on the method used and the specific experimental conditions. Microinjection has shown high success rates in various aquatic species, including shrimp, due to its precision and control over the amount of genetic material delivered (Abdelrahman et al., 2021; Lane et al., 2021; Gultom, 2023). Electroporation and lipofection offer alternative approaches with varying degrees of success, depending on the optimization of protocols and reagents used. Viral vectors, while promising, require further research to ensure their safety and effectiveness in shrimp. 3.4 Case study: successful gene transfer using CRISPR-Cas9 in shrimp A notable example of successful gene transfer in shrimp involves the use of CRISPR-Cas9 technology. This method has revolutionized genetic engineering by allowing precise and targeted modifications of the genome. In a study involving zebrafish, automated microinjection of CRISPR/Cas9 components demonstrated high efficiency and success rates, highlighting the potential for similar applications in shrimp (Cordero-Maldonado et al., 2019). The use of CRISPR-Cas9 in shrimp can facilitate the study of gene functions and the development of genetically improved shrimp strains, contributing to advancements in aquaculture and biotechnology. 4 Cellular Applications of Gene Transfer 4.1 Gene editing for disease resistance Gene editing technologies, such as CRISPR/Cas9, have been instrumental in enhancing disease resistance in shrimp. By integrating antimicrobial peptide genes (AMGs) through genome editing, researchers have successfully modulated the innate immune system of aquatic animals, leading to improved survival rates and immune responses against pathogens. This approach has shown promise in reducing bacterial colony-forming units and increasing lysozyme activity, which are critical for disease resistance. Additionally, the expression of immune-related genes such as IL, IKβ, TGFβ, C3b, and TLR is significantly enhanced, contributing to a robust immune response (Wang and Cheng, 2023). 4.2 Enhancing growth and reproduction Gene transfer techniques have also been employed to enhance growth and reproduction in shrimp. For instance, dietary supplementation with specific nutrients, such as hydrolyzed yeast and Bacillus licheniformis, has been shown to improve feed efficiency and protein efficiency ratios in shrimp. These dietary interventions, facilitated by gene transfer, lead to better growth performance and reproductive outcomes. Moreover, the upregulation of genes related to growth and immune responses, such as CAT, GPX, SOD, Pen-3a, and PPO, further supports the enhanced growth and reproductive capabilities of shrimp (Chen et al., 2020).
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