Animal Molecular Breeding, 2025, Vol.15, No.2, 49-59 http://animalscipublisher.com/index.php/amb 54 suitable tools, such as CRISPR/Cas9 or ISDra2-TnpB, and predict their editing efficiency and accuracy (Zhang et al., 2019; Farooq et al., 2024). Doing so can arrange experiments more scientifically, such as conducting gene knockout and knock-in simultaneously, to improve some specific traits, such as making goats more disease-resistant, or enabling them to produce more of a certain protein (Figure 3) (Zhang et al., 2018; Feng et al., 2024; Li et al., 2024). Figure 3 Cas9-mediated HNP1 knock-in in goats at the CSN2locus (Adopted from Li et al., 2024) Image caption: (a) Schematic diagram of experiment design for HNP1 insertion into goat CSN2. sgRNA left and right were designed to target exon 7 of CSN2. The plasmid PUC57-HNP1 with the left arm (LA), 2A-HNP1, and the right arm (RA) was used as a homologous repair template for HNP1 insertion. Four primer sets were utilized for genome editing detection. In wild type (WT) and CSN2 KO goats (repaired through the NHEJ method), only a 420 bp fragment could be amplified using the T7E1 primer sets. In HNP1-inserted goats (repaired through HDR method), an additional 813 bp fragment could be amplified. The other three primer sets, HDR_LA, HDR_RA, and HNP1, only amplified the expected fragments in HNP1-inserted goats. (b) PCR analysis using the four primer sets. (c) The two goats (H2 and P2) with HNP1 insertion at CSN2 locus. (d) TA clone sequencing results of HDR_LA and HDR_RA amplicons. * following the T2A-HNP1, indicating the stop codon (Adopted from Li et al., 2024) 7 Case Study: AI-Assisted Genomic Selection in Dairy Goat Breeding 7.1 Background and breeding objectives The breeding of dairy goats is mainly aimed at enhancing economic traits such as milk production and milk protein, as well as body shape characteristics that affect the lifespan and production efficiency of goats. Most traditional methods rely on pedigree and expression data for seed selection. However, if the heritability of some
RkJQdWJsaXNoZXIy MjQ4ODYzNA==