Shin Sung Wook, Kim Su Hyun, Gasselin Aghiles, Lee Gyun Min, Lee Jae Seong
Department of Molecular Science and Technology, Ajou University, Suwon, 16499, Republic of Korea.
Department of Biological Sciences, KAIST, Daejeon, 34141, Republic of Korea.
Sci Data. 2025 Jan 15;12(1):71. doi: 10.1038/s41597-025-04438-6.
Chinese hamster ovary (CHO) cells play a pivotal role in the production of recombinant therapeutics. In the present study, we conducted a genome-scale pooled CRISPR knockout (KO) screening using a virus-free, recombinase-mediated cassette exchange-based platform in CHO-K1 host and CHO-K1 derived recombinant cells. Genome-wide guide RNA (gRNA) amplicon sequencing data were generated from cell libraries, as well as short- and long-term KO libraries, and validated through phenotypic assessment and gRNA read count distribution. Additionally, we obtained gRNA amplicon sequencing data from the highly productive recombinant cell populations. By analyzing these datasets, essential genes involved in cell fitness as well as functional target genes associated with therapeutic protein production can be identified. Collectively, our next-generation sequencing datasets, derived from a robust and reliable CRISPR screening method, provide valuable insights into CHO genomic functions, advancing the development of next-generation CHO factories.
中国仓鼠卵巢（CHO）细胞在重组治疗药物的生产中起着关键作用。在本研究中，我们使用基于无病毒、重组酶介导的盒式交换平台，在CHO-K1宿主细胞和CHO-K1衍生的重组细胞中进行了全基因组规模的混合CRISPR基因敲除（KO）筛选。从细胞文库以及短期和长期KO文库中生成全基因组引导RNA（gRNA）扩增子测序数据，并通过表型评估和gRNA读数计数分布进行验证。此外，我们从高产重组细胞群体中获得了gRNA扩增子测序数据。通过分析这些数据集，可以鉴定参与细胞适应性的必需基因以及与治疗性蛋白质生产相关的功能靶基因。总的来说，我们从强大且可靠的CRISPR筛选方法中获得的下一代测序数据集，为CHO基因组功能提供了有价值的见解，推动了下一代CHO工厂的发展。
