Kaczmarczyk Lech, Jackson Walker S
German Centre for Neurodegenerative Diseases (DZNE), Bonn, German.
Swiss Med Wkly. 2015 Oct 29;145:w14186. doi: 10.4414/smw.2015.14186. eCollection 2015.
The humble house mouse has long been a workhorse model system in biomedical research. The technology for introducing site-specific genome modifications led to Nobel Prizes for its pioneers and opened a new era of mouse genetics. However, this technology was very time-consuming and technically demanding. As a result, many investigators continued to employ easier genome manipulation methods, though resulting models can suffer from overlooked or underestimated consequences. Another breakthrough, invaluable for the molecular dissection of disease mechanisms, was the invention of high-throughput methods to measure the expression of a plethora of genes in parallel. However, the use of samples containing material from multiple cell types could obfuscate data, and thus interpretations. In this review we highlight some important issues in experimental approaches using mouse models for biomedical research. We then discuss recent technological advances in mouse genetics that are revolutionising human disease research. Mouse genomes are now easily manipulated at precise locations thanks to guided endonucleases, such as transcription activator-like effector nucleases (TALENs) or the CRISPR/Cas9 system, both also having the potential to turn the dream of human gene therapy into reality. Newly developed methods of cell type-specific isolation of transcriptomes from crude tissue homogenates, followed by detection with next generation sequencing (NGS), are vastly improving gene regulation studies. Taken together, these amazing tools simplify the creation of much more accurate mouse models of human disease, and enable the extraction of hitherto unobtainable data.
普通家鼠长期以来一直是生物医学研究中常用的模型系统。引入位点特异性基因组修饰的技术为其先驱者带来了诺贝尔奖，并开启了小鼠遗传学的新时代。然而，这项技术非常耗时且技术要求很高。因此，许多研究人员继续采用更简便的基因组操作方法，尽管由此产生的模型可能存在被忽视或低估的后果。另一项对疾病机制的分子剖析具有重要价值的突破是发明了高通量方法，能够并行测量大量基因的表达。然而，使用包含多种细胞类型材料的样本可能会使数据模糊不清，从而影响解释。在这篇综述中，我们重点介绍了使用小鼠模型进行生物医学研究的实验方法中的一些重要问题。然后，我们讨论了小鼠遗传学领域的最新技术进展，这些进展正在彻底改变人类疾病研究。由于有了引导性核酸内切酶，如转录激活样效应核酸酶（TALENs）或CRISPR/Cas9系统，现在可以轻松地在精确位置对小鼠基因组进行操作，这两者也都有可能将人类基因治疗的梦想变为现实。新开发的从粗组织匀浆中进行细胞类型特异性转录组分离的方法，随后通过下一代测序（NGS）进行检测，正在极大地改进基因调控研究。综上所述，这些惊人的工具简化了更精确的人类疾病小鼠模型的创建，并能够获取迄今无法获得的数据。
