Upstream Research and Development, MilliporeSigma, St. Louis, Missouri, USA.
Bioinformatics, IT R&D Applications, Merck (Sigma-Aldrich Chemicals Pvt. Ltd., A subsidiary of Merck KGaA, Darmstadt, Germany), Bangalore, India.
Biotechnol Bioeng. 2022 Dec;119(12):3632-3646. doi: 10.1002/bit.28226. Epub 2022 Sep 23.
Chinese hamster ovary (CHO) cells have been used as the industry standard for the production of therapeutic monoclonal antibodies for several decades. Despite significant improvements in commercial-scale production processes and media, the CHO cell has remained largely unchanged. Due to the cost and complexity of whole-genome sequencing and gene-editing it has been difficult to obtain the tools necessary to improve the CHO cell line. With the advent of next-generation sequencing and the discovery of the CRISPR/Cas9 system it has become more cost effective to sequence and manipulate the CHO genome. Here, we provide a comprehensive de novo assembly and annotation of the CHO-K1 based CHOZN® GS genome. Using this platform, we designed, built, and confirmed the functionality of a whole genome CRISPR guide RNA library that will allow the bioprocessing community to design a more robust CHO cell line leading to the production of life saving medications in a more cost-effective manner.
中国仓鼠卵巢 (CHO) 细胞已被用作生产治疗性单克隆抗体的行业标准数十年。尽管商业规模的生产工艺和培养基有了显著的改进,但 CHO 细胞基本上保持不变。由于全基因组测序和基因编辑的成本和复杂性,获得改进 CHO 细胞系所需的工具一直具有挑战性。随着下一代测序技术的出现和 CRISPR/Cas9 系统的发现,对 CHO 基因组进行测序和操作变得更加经济实惠。在这里,我们提供了基于 CHO-K1 的 CHOZN® GS 基因组的全面从头组装和注释。使用这个平台,我们设计、构建并验证了一个全基因组 CRISPR 向导 RNA 文库的功能,该文库将使生物加工界能够设计出更稳健的 CHO 细胞系,以更具成本效益的方式生产挽救生命的药物。