Animal Molecular Breeding 2024, Vol.14, No.1, 1-9 http://animalscipublisher.com/index.php/amb 3 Figure 2 Gene editing technology 1.2 Methods and techniques for gene editing in crucian carp embryos Editing genes in crucian carp embryos is a complex and precise process that requires carefully designed and tightly controlled operational steps. The design of guide RNA is the starting point for crucian carp gene editing. Guide RNA must highly specifically match the DNA sequence of the target gene, thus requiring in-depth genomic knowledge and the support from computational biology tools. The preparation of guide RNA and Cas9 protein must be carried out in the laboratory. This requires high-purity and highly active complexes to ensure their effectiveness inside embryos. The prepared CRISPR-Cas9 complex must be precisely injected into crucian carp embryos. This is an operation that requires high technical skills and accuracy to ensure the successful introduction of the complex into the embryos. Subsequent verification of successful editing is necessary to ensure the desired genetic changes have occurred. This typically involves extracting DNA from edited embryos and conducting gene sequencing to confirm the editing effects (Liu et al., 2004). In crucian carp embryo gene editing, selecting the appropriate target genes is crucial. Typically, genes closely associated with growth rate and fat metabolism are chosen as editing targets. Selecting key genes that control crucian carp growth rate and fat metabolism requires in-depth genetic and physiological research. This may involve a comprehensive analysis of existing literature and experimental validation. Additionally, special attention must be paid to the design of guide RNA to ensure its complete match with the target gene's DNA sequence. This requires precise design and analysis of the guide RNA sequence. By gaining a deep understanding of the CRISPR-Cas9 system and the methods and techniques of gene editing in crucian carp embryos, this study provides a scientific basis for applying gene editing to improve carp quality. These key technologies and steps contribute essential theoretical and practical foundations to the study of gene editing in crucian carp embryos. 2 Growth Rate Control Mechanisms 2.1 Gene regulation of growth hormones and growth factor The growth rate of crucian carp is complex influenced by various biological processes and gene regulations. Among them, growth hormones and growth factors playing a crucial role in the control of fish growth speed. Growth hormone (GH) is a multifunctional protein hormone that plays a significant role in the growth of crucian carp. The pituitary gland secretes growth hormone, which initiates a series of complex signal transduction pathways by binding to the growth hormone receptor (GHR). These pathways activate cell division and growth, leading to the rapid development of the fish. Through gene editing techniques, it is possible to precisely control the expression level of the GHR gene, enhancing or attenuating the signal transduction of growth hormones,
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