使用DNA修复基因表达的CHO细胞系稳定性的预测
Lauren T Cordova1, Hussain Dahodwala1,2, Rebecca Cooley3
1Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
像Lig4和Xrcc6这样的DNA修复基因可能会预测中国仓鼠卵巢 (CHO) 细胞系的稳定性. 老细胞表达率较低表明潜在的生产不稳定性,有助于生物制药制造.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
背景情况:
- 中国仓鼠卵巢 (CHO) 细胞对于生物制药生产至关重要.
- 生产不稳定,生产率随着时间的推移而下降,是行业面临的主要挑战.
- 预测细胞系稳定性可以通过早期识别强大的克隆来优化制造.
研究的目的:
- 研究DNA修复基因作为预测CHO细胞系稳定性的潜在生物标志物.
- 为了将早期的DNA修复基因表达与长期生产稳定性相关联.
- 为了确定与CHO-DG44细胞系不稳定性相关的特定基因.
主要方法:
- 在40多个CHO细胞系中评估了5个DNA修复基因 (Fam35a,Lig4,Palb2,Pari,Xrcc6) 的表达.
- 早期通行时相关的基因表达 (<30种人口翻倍水平) 在晚期通行时具有稳定性 (60+ PDL).
- 利用主要成分分析来识别区分稳定和不稳定细胞系的标记物.
主要成果:
- 两个基因,Lig4和Xrcc6,在不稳定的CHO-DG44细胞系中表达更高.
- 副本数量丢失被确定为导致这些线路不稳定的机制.
- 较低的DNA修复基因表达与所有细胞系的细胞年龄增加相关.
结论:
- 修复DNA基因表达提供了对CHO细胞长期行为的见解.
- 在特定情况下,Lig4和Xrcc6表达水平显示出预测细胞系稳定性的潜力.
- 这项研究可能会导致生物制药生产中更有效地选细胞系.
更多相关视频
06:59Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
Published on: March 31, 2022
08:53Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1
Published on: February 17, 2011
相关概念视频
DNA Damage can Stall the Cell Cycle
DNA Damage Can Stall the Cell Cycle
Nucleotide Excision Repair
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
Overview of DNA Repair
Chemically...
Fixing Double-strand Breaks
Base-pairing and DNA Repair
