Department of Microbiology and Immunology, Institute of Marine and Environmental Technology, University of Maryland Baltimore, Baltimore, MD, USA.
State Key Laboratory for Biocontrol, School of Marine Sciences, Sun Yat-Sen University, Guangzhou, People's Republic of China.
Methods Mol Biol. 2022;2442:425-443. doi: 10.1007/978-1-0716-2055-7_23.
Techniques for disrupting gene expression are invaluable tools for the analysis of the biological role of a gene product. Because of its genetic tractability and multiple advantages over conventional mammalian models, the zebrafish (Danio rerio) is recognized as a powerful system for gaining new insight into diverse aspects of human health and disease. Among the multiple mammalian gene families for which the zebrafish has shown promise as an invaluable model for functional studies, the galectins have attracted great interest due to their participation in early development, regulation of immune homeostasis, and recognition of microbial pathogens. Galectins are β-galactosyl-binding lectins with a characteristic sequence motif in their carbohydrate recognition domains (CRDs), that constitute an evolutionary conserved family ubiquitous in eukaryotic taxa. Galectins are emerging as key players in the modulation of many important pathological processes, which include acute and chronic inflammatory diseases, autoimmunity and cancer, thus making them potential molecular targets for innovative drug discovery. Here, we provide a review of the current methods available for the manipulation of gene expression in the zebrafish, with a focus on gene knockdown [morpholino (MO)-derived antisense oligonucleotides] and knockout (CRISPR-Cas) technologies.
基因表达干扰技术是分析基因产物生物学功能的宝贵工具。由于其遗传易操作性以及相对于传统哺乳动物模型的多种优势,斑马鱼(Danio rerio)被认为是深入了解人类健康和疾病各个方面的强大系统。在多种哺乳动物基因家族中,斑马鱼作为功能研究的宝贵模型显示出巨大的潜力,其中半乳糖凝集素因其参与早期发育、免疫稳态调节以及识别微生物病原体而引起了极大的关注。半乳糖凝集素是β-半乳糖结合凝集素,其碳水化合物识别结构域(CRD)中有一个特征序列基序,构成了在真核生物分类中普遍存在的进化保守家族。半乳糖凝集素正在成为许多重要病理过程(包括急性和慢性炎症性疾病、自身免疫和癌症)调节的关键参与者,因此成为创新药物发现的潜在分子靶标。在这里,我们综述了目前在斑马鱼中操纵基因表达的可用方法,重点介绍基因敲低[基于 morpholino(MO)的反义寡核苷酸]和基因敲除(CRISPR-Cas)技术。