Dimitrakopoulou Dionysia, Tulkens Dieter, Van Vlierberghe Pieter, Vleminckx Kris
Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
Cancer Research Institute Ghent (CRIG), Ghent, Belgium.
Front Physiol. 2019 Feb 1;10:48. doi: 10.3389/fphys.2019.00048. eCollection 2019.
Aquatic vertebrate organisms such as zebrafish have been used for over a decade to model different types of human cancer, including hematologic malignancies. However, the introduction of gene editing techniques such as CRISPR/Cas9 and TALEN, have now opened the road for other organisms featuring large externally developing embryos that are easily accessible. Thanks to its unique diploid genome that shows a high degree of synteny to the human, combined with its relatively short live cycle, has now emerged as an additional powerful aquatic model for studying human disease genes. Genome editing techniques are very simple and extremely efficient, permitting the fast and cheap generation of genetic models for human disease. Mosaic disruption of tumor suppressor genes allows the generation of highly penetrant and low latency cancer models. While models for solid human tumors have been recently generated, genetic models for hematologic malignancies are currently lacking for Here we describe our experimental pipeline, based on mosaic genome editing by CRISPR/Cas9, to generate innovative and high-performing leukemia models in . These add to the existing models in zebrafish and will extend the experimental platform available in aquatic vertebrate organisms to contribute to the field of hematologic malignancies. This will extend our knowledge in the etiology of this cancer and assist the identification of molecular targets for therapeutic intervention.
诸如斑马鱼之类的水生脊椎动物已被用于模拟包括血液系统恶性肿瘤在内的不同类型的人类癌症长达十多年。然而,诸如CRISPR/Cas9和TALEN等基因编辑技术的引入,现在为其他具有易于获取的大型外部发育胚胎的生物开辟了道路。由于其独特的二倍体基因组与人类基因组具有高度的同线性,再加上其相对较短的生命周期,它现在已成为研究人类疾病基因的另一种强大的水生模型。基因组编辑技术非常简单且极其高效,能够快速且低成本地生成人类疾病的遗传模型。肿瘤抑制基因的镶嵌式破坏能够生成高侵袭性和低潜伏期的癌症模型。虽然最近已经生成了人类实体瘤模型,但目前缺乏血液系统恶性肿瘤的遗传模型。在此,我们描述了基于CRISPR/Cas9镶嵌式基因组编辑的实验流程,以在斑马鱼中生成创新的、高性能的白血病模型。这些模型补充了斑马鱼现有的模型,并将扩展水生脊椎动物可用的实验平台,为血液系统恶性肿瘤领域做出贡献。这将扩展我们对这种癌症病因的认识,并有助于确定治疗干预的分子靶点。