De novo assembly and annotation of the CHOZN® GS−/− genome supports high‐throughput genome‐scale screening

• Published in: Biotechnology and bioengineering Vol. 119; no. 12; pp. 3632 - 3646
• Main Authors: Kretzmer, Corey, Narasimhan, Rajagopalan Lakshmi, Lal, Rahul Deva, Balassi, Vincent, Ravellette, James, Kotekar Manjunath, Ajaya Kumar, Koshy, Jesvin Joy, Viano, Marta, Torre, Serena, Zanda, Valeria M., Kumravat, Mausam, Saldanha, Keith Metelo Raul, Chandranpillai, Harikrishnan, Nihad, Ifra, Zhong, Fei, Sun, Yi, Gustin, Jason, Borgschulte, Trissa, Liu, Jiajian, Razafsky, David
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-12-2022; John Wiley and Sons Inc
• Subjects: Animals; Annotations; Assembly; Bioprocessing; cell line engineering; Chinese hamster ovary; CHO Cells; Cricetinae; Cricetulus; CRISPR; CRISPR-Cas Systems - genetics; Gene sequencing; Genome - genetics; Genomes; genome‐wide pooled screens; Industry standards; Monoclonal antibodies; Nucleotide sequence; RNA, Guide, CRISPR-Cas Systems - genetics; CRISPR; cell line engineering; Bioprocessing; Chinese hamster ovary; genome-wide pooled screens
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.28226

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.