CHO细胞基因组中稳定hot spot位点研究进展

Research Progress on Stable Hot Spot Sites in the Genome of CHO Cells

近些年来,治疗性重组蛋白类药物是生物制药领域研究的热点。工业化生产中常用于重组蛋白表达的细胞系是中国仓鼠卵巢(Chinese hamster ovary,CHO)细胞。传统CHO细胞系的表达大多数基于随机整合的方式,这可能会使目标基因整合到异染色质区域或者不稳定的染色质区域,导致CHO细胞表达不稳定,需要多轮筛选才能获得理想的表达细胞系。最新研究表明,外源基因在CHO细胞预测/特定的基因组位点中进行特异性整合,可以使重组CHO细胞的表达保持长期一致性和稳定性。CHO细胞基因组中高效稳定的转录整合位点被称为热点(hot spot)。阐述CHO细胞基因组稳定的hot spot位点近几年的研究进展,其中包括热门的hot spot位点,以及如何研究新的hot spot位点的方法。总结如何将外源基因高效定位于预测的CHO细胞hot spot位点,实现高水平稳定的表达重组蛋白,为发现新的有效的hot spot位点,构建稳定表达CHO细胞系提供参考。

In recent years,therapeutic recombinant protein drugs have become a research hot topic in the field of biopharmaceuticals research.Chinese hamster ovary cell lines are commonly used for recombinant protein expression in industrial production.The traditional expression of CHO cell lines is mostly based on random integration.However,random integration may result in target genes targeted into heterochromatin regions or unstable chromatin regions,causing unstable expression in CHO cell lines,and in order to obtain the ideal expression cell line researchers have to do multiple rounds of screening.The latest research proves that GOI integrates specifically in the prediction or specific genomic sites of CHO cells,which can maintain long-term consistency and stability in the expression of recombinant CHO cells.The integration sites for efficient and stable transcription in the genome of CHO cells are called hot spots.This review describes the latest research progress of stable hot spot sites in the CHO cell genome,summarizes the research methods of these sites,and concludes how to locate the GOI at predicted gene locus efficiently,to achieve high-level and stable expression recombinant protein,and to help researchers discover new or effective hot spot sites and then construct stable expression CHO cell lines.