Expanding the CRISPR Toolbox for Chinese Hamster Ovary Cell Line Engineering
Abstract
Recombinant biotherapeutic proteins are used in the treatment of some of the most serious diseases, such as cancer and autoimmune diseases, affecting millions of people worldwide. In this field, Chinese hamster ovary (CHO) cells dominate as the expression host of choice. For decades CHO cells have delivered high quality products approved by regulatory agencies, yet the underlying mechanisms governing their behaviour are not well understood. To meet the increasing global demand from a growing and aging population it is imperative to be able to precisely manipulate cells to obtain desirable traits. Recent advances in unveiling the genetic context and developments in genetic engineering tools have unlocked a new era for understanding and changing the nature of CHO cells. The gene editing technology CRISPR has gained foothold as a powerful genetic engineering system in multiple mammalian systems and has proven useful for engineering CHO cells. We here present the application of novel CRISPR techniques in CHO cells with the aim of expanding the tool box to enable more diverse CHO cell line engineering and target discovery. We focus on three methods: (1) CRISPR interference to repress transcription, (2) CRISPR activation to enhance transcription, and (3) large-scale CRISPR knockout screening to enable high-throughput novel target identification. We conclude that these three tools can be used individually or in conjunction to permit a multilevel exploration of the CHO phenotypic space, which should permit more rapid development of rationally engineered CHO cell lines for biotherapeutic protein production.
Info
Thesis PhD, 2019
UN SDG Classification
DK Main Research Area
Science/Technology
