Advanced Cell Line Development with Cas-CLOVER and Transposases
Explore Demeetra's technologies for cell line development, Cas-CLOVER, a precise alternative to CRISPR/Cas9, and transposases for stable protein production in pharmaceutical bioprocessing
Optimizing Cell Lines for Efficient and Scalable Bioprocessing
Efficient cell line development and upstream bioprocessing are essential for producing biotherapeutics and other critical proteins at commercial scale, ensuring high efficiency, cost-effectiveness, and consistent product quality.
Leveraging advanced tools like Cas-CLOVER for precise knockouts and knock-ins, alongside transposase technologies like piggyBac for stable integration at highly expressed genomic sites, streamlines the optimization of mammalian expression platforms such as CHO and HEK293 cells. These innovations enable faster, more reliable, and scalable production of life-saving biotherapeutics.
Cas-CLOVER, Larger Knock-ins and More Precise than CRISPR/Cas9
Cas-CLOVER offers a transformative approach to cell line development with its clean dimeric nuclease structure minimizing off-target effects while leveraging staggered cuts, increasing the size of deletions and enhancing the efficiency of knock-in events. Unlike traditional CRISPR/Cas9 systems, which have been limited to knock-ins of 7 kilobases, Cas-CLOVER can seamlessly integrate sequences of at least 20 kilobases at precise genomic locations. These capabilities make it an ideal tool for platform enhancement and applications requiring the insertion of complex genetic constructs such as next generation biotherapeutics.
Cas-CLOVER Validated in Cell Engineering
The Cas-CLOVER technology has been tested in CHO and HEK293 cells. In-house comparisons of Cas-CLOVER with Cas9 in CHO cells showed either superior or comparable editing efficiencies. Additionally, glutamine synthetase (GS) was knocked out in CHO cells, with KO success evidenced by reduced viability and inhibited cell growth in glutamine-deprived media.
Besides targeted gene inactivation, Cas-CLOVER is more efficient than CRISPR/Cas9 at generating targeted knock-ins at any genomic locus, including "hotspot" sites, for consistent protein expression.
PiggyBac For Stable Protein Production
PiggyBac is a DNA transposase/transposon system that introduces genetic cargo stably into the genome for consistently high protein production.
PiggyBac Improves Protein Production Compared to Conventional Plasmid Expression
PiggyBac has been validated for antibody production in both CHO cell pools and single cell clones, as shown by Eli Lilly and others. Protein production notably increased without affecting cell growth or viability. Additionally, protein expression remained stable over 60 generations in CHO cells.
See How Scientists Are Using Our Gene Editing Tools
Elanco Research Scientist Kayla Bean, Ph.D, explored how Cas-CLOVER optimizes CHO cells for cell line development.
Kayla Bean, Ph.D.
Research Scientist, Discovery Research
Elanco Animal Health
“The flexibility of the guide RNA design makes the system easy to use and gives high specificity due to the use of the two guide RNAs.
It is very efficient due to the ability of the dead Cas9s [dCas's] to recognize the current area of DNA, and since the clo51 nuclease can only cut when dimerized, the system has high fidelity.”
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