CRISPR and other gene editing tools are revolutionizing cell line engineering due to the speed and ease of performing edits. They are a cheaper, more efficient, targeted and easier way to modify the gene. This whitepaper explores in depth ways that CRISPR and gene editing tools are being used in cell line development and engineering. Read the summary below or download the free whitepaper here.
Biopharmaceuticals pipelines using mammalian cells still bear with cellular and platform limitations compared to bacterial or yeast-based expression systems. How can we overcome diminished growth, low yield, and stress vulnerabilities?
While the role of biologics in treating human diseases has evolved dramatically over the past decade, so has the technology to engineer cell factories. In fact, rational genetic engineering of cell factories to enhance biotherapeutic proteins has become a reality catalyzed by the publication of the genome sequences of multiple CHO cell lines and the Chinese hamster’s.
Novel “designer CHO cells” which modulate PTMs of recombinant proteins by genome editing is today a reality. It is now possible single editing to multiple genes for knocking-in or knocking-out the genome of yeast and mammalian cells, with “surgical accuracy” (one DNA base pair precision) quickly, in an efficient manner. Eventually, allowing the development of a cost-effective recombinant therapeutic protein.
Notably, the bacterial clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing tool has drastically improved the genome-editing—making it faster, easier, cheaper, and more efficient. Remarkably, this system needs only one protein and one RNA molecule to achieve RNA-programmed DNA cleavage. Indeed, that has enabled researchers in the CHO community to elucidate the mechanistic basis behind the high-level production of proteins and product quality attributes of interest. Presently, CHO gene editing emphasis is being directed toward expanding the product diversity, controlling and improving product quality and yields. While routine PTM optimization across the cell's glycosylation machinery is not there yet, combining the expression from nonmammalian hosts with human-like antibody glycosylation performed in engineered yeast or even plants would be achievable soon.
Sooner, the potential of promoter engineering to achieve precision transcriptional control for CHO cell synthetic biotechnology would be a reality. Similarly, multiplexed genome editing could further allow the examination and manipulation of whole genomes or protein networks.
“Multiplexed” editing constituted a breakthrough that has allowed to generate knock-out and knock-in clones, for metabolic engineering of mammalian cells and yeasts. Certainly, it has enabled concurrent consistent, stable and high expression of a complex recombinant therapeutic protein such as monoclonal antibodies (mAbs).
As summarized by Doudna and Barrangou (two of the creators of the system), the potential for CRISPR applications is huge and will “affect almost every aspect of life, and provide inspiration for future technological breakthroughs.”
Are there constraints for commercial human therapeutic applications of gene editing, given the current IP landscape? How important is the outcome of the patent interference (UC – U of Vienna vs. “Boston Biotech Cluster”) to the future of gene editing?
Are there uncertainties in translational research and clinical trials due to broad exclusive licenses over CRISPR technologies IP? Is CRISPR genome-editing know-how going to define future applications and further developments of it as nonobvious? Will commercial sub-licenses be needed for follow-on applications of CRISPR-Cas technology? If so, when is this going to happen?