位点特异性整合开启了精确CHO细胞系工程的新时代。
Site-specific Integration Ushers in a New Era of Precise CHO Cell Line Engineering.
作者信息
Hamaker Nathaniel K, Lee Kelvin H
机构信息
Delaware Biotechnology Institute, Newark, DE.
Chemical and Biomolecular Engineering, University of Delaware, Newark, DE.
出版信息
Curr Opin Chem Eng. 2018 Dec;22:152-160. doi: 10.1016/j.coche.2018.09.011. Epub 2018 Oct 23.
Abstract
Chinese hamster ovary (CHO) cells are widely used for the production of therapeutic proteins. Customarily, CHO production cell lines are established through random integration, which requires laborious screening of many clones to isolate suitable producers. In contrast, site-specific integration (SSI) accelerates cell line development by targeting integration of transgenes to pre-validated genomic loci capable of supporting high and stable expression. To date, a relatively small number of these so called 'hot spots' have been identified, mainly through empirical methods. Nevertheless, nuclease-mediated and recombinase-mediated SSI have revolutionized cell line engineering by enabling rational and reproducible transgene targeting.
摘要
中国仓鼠卵巢（CHO）细胞被广泛用于治疗性蛋白质的生产。通常，CHO生产细胞系是通过随机整合建立的，这需要对许多克隆进行费力的筛选以分离出合适的生产者。相比之下，位点特异性整合（SSI）通过将转基因靶向整合到能够支持高表达和稳定表达的预先验证的基因组位点来加速细胞系的开发。迄今为止，主要通过经验方法鉴定出了相对较少的这些所谓的“热点”。然而，核酸酶介导的和重组酶介导的SSI通过实现合理且可重复的转基因靶向，彻底改变了细胞系工程。
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