Feng Chiguang, Nita-Lazar Mihai, González-Montalbán Nuria, Wang Jingyu, Mancini Justin, Ravindran Chinnarajan, Ahmed Hafiz, Vasta Gerardo R
Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, MD, USA.
Methods Mol Biol. 2015;1207:327-41. doi: 10.1007/978-1-4939-1396-1_22.
Techniques for disrupting gene expression are invaluable tools for the analysis of the biological role(s) 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), which comprise 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.
破坏基因表达的技术是分析基因产物生物学作用的宝贵工具。由于斑马鱼（Danio rerio）具有遗传易处理性且相对于传统哺乳动物模型具有多种优势，它被公认为是一个强大的系统，有助于深入了解人类健康和疾病的各个方面。在多个哺乳动物基因家族中，斑马鱼已显示出作为功能研究的宝贵模型的潜力，其中半乳糖凝集素因其参与早期发育、免疫稳态调节以及对微生物病原体的识别而备受关注。半乳糖凝集素是β-半乳糖苷结合凝集素，在其碳水化合物识别结构域（CRD）中具有特征性序列基序，该结构域构成了一个在真核生物分类群中普遍存在的进化保守家族。半乳糖凝集素正逐渐成为许多重要病理过程调节中的关键参与者，这些病理过程包括急性和慢性炎症性疾病、自身免疫和癌症，因此使其成为创新药物发现的潜在分子靶点。在这里，我们综述了目前可用于在斑马鱼中操纵基因表达的方法，重点是基因敲低[吗啉代（MO）衍生的反义寡核苷酸]和基因敲除（CRISPR-Cas）技术。
