Please use this identifier to cite or link to this item: https://scholarbank.nus.edu.sg/handle/10635/212685
Title: DEVELOPMENT OF NOVEL EXPRESSION SYSTEMS FOR RELIABLE GENETIC ENGINEERING IN CHINESE HAMSTER OVARY CELLS
Authors: SUN TAO
Keywords: CHO cells, site-specific integration, CRISPR-Cas9, proline, selection, monoclonal antibody
Issue Date: 5-Aug-2019
Citation: SUN TAO (2019-08-05). DEVELOPMENT OF NOVEL EXPRESSION SYSTEMS FOR RELIABLE GENETIC ENGINEERING IN CHINESE HAMSTER OVARY CELLS. ScholarBank@NUS Repository.
Abstract: Chinese hamster ovary (CHO) cells are the dominant mammalian cell lines for biologics. However, the CHO cell line development for biologics production remains time and labour consuming. Herein we describe novel expression systems for CHO cells to be applicable for genetic engineering via CRISPR-Cas9. The main issues in conventional integration of FRT site into the genome are addressed by improving transfected DNA integration through a hybrid FLP-FRT recombination and CRISPR-Cas9. In addition, available selection system for CHO cells were expanded based on proline metabolism and this antibiotic-free selection could be applicable for a variety of biologics production. By expressing fluorescent proteins and a monoclonal antibody, it demonstrated that platform cell lines and antibiotic-free selection systems were reliable and efficient. Considering the scientific innovation and high potential in a number of application areas, this achievement would be of great potential in industrial application.
URI: https://scholarbank.nus.edu.sg/handle/10635/212685
Appears in Collections:Ph.D Theses (Open)

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