Plazyo Olesya, Hao Weilong, Jin Jian-Ping
Department of Physiology, School of Medicine, Wayne State University, Detroit, MI, United States.
Department of Biological Sciences, Wayne State University, Detroit, MI, United States.
Front Bioeng Biotechnol. 2020 Feb 28;8:42. doi: 10.3389/fbioe.2020.00042. eCollection 2020.
While the rapid development of CRISPR/CAS9 technology has allowed for readily performing site-specific genomic editing in non-rodent species, an emerging challenge is to select the most suitable species to generate animal models for the study of human biology and diseases. Improving CRISPR/CAS9 methodology for more effective and precise editing in the rabbit genome to replicate human disease is an active area of biomedical research. Although rabbits are more closely related to humans than mice (based on DNA sequence analysis), our whole-genome protein database search revealed that rabbits have more missing human protein sequences than mice. Hence, precisely replicating human diseases in rabbits requires further consideration, especially in studies involving essential functions of the missing proteins. For example, rabbits lack calponin 2, an actin-associated cytoskeletal protein that is important in the pathogenesis of inflammatory arthritis, atherosclerosis, and calcific aortic valve disease. The justification of using rabbits as models for human biomedical research is based on their larger size and their closer phylogenetic distance to humans (based on sequence similarity of conserved genes), but this may be misleading. Our findings, which consider whole-genome protein profiling together with actual protein expressions, serve as a warning to the scientific community to consider overall conservation as well as the conservation of specific proteins when choosing an animal model to study a particular aspect of human biology prior to investing in genetic engineering.
虽然CRISPR/CAS9技术的迅速发展使得在非啮齿类动物中轻松进行位点特异性基因组编辑成为可能,但一个新出现的挑战是选择最合适的物种来生成用于研究人类生物学和疾病的动物模型。改进CRISPR/CAS9方法以在兔基因组中进行更有效和精确的编辑以复制人类疾病,是生物医学研究的一个活跃领域。尽管基于DNA序列分析,兔子比小鼠与人类的亲缘关系更近,但我们对全基因组蛋白质数据库的搜索显示,兔子缺失的人类蛋白质序列比小鼠更多。因此,在兔子中精确复制人类疾病需要进一步考虑,特别是在涉及缺失蛋白质基本功能的研究中。例如,兔子缺乏钙调蛋白2,这是一种与肌动蛋白相关的细胞骨架蛋白,在炎症性关节炎、动脉粥样硬化和钙化性主动脉瓣疾病的发病机制中很重要。将兔子用作人类生物医学研究模型的理由基于它们更大的体型以及它们与人类更接近的系统发育距离(基于保守基因的序列相似性),但这可能会产生误导。我们的研究结果,将全基因组蛋白质分析与实际蛋白质表达结合起来考虑,向科学界发出了一个警告,即在投资于基因工程之前,选择动物模型来研究人类生物学的特定方面时,要考虑整体保守性以及特定蛋白质的保守性。