CleanCut GS CHO
Glutamine synthetase (GS) selection is the industry standard for the efficient identification of high-titer CHO cell pools or clones for recombinant therapeutic protein production. Demeetra's CleanCut GS knockout suspension cells are engineered with the proprietary Cas-CLOVER targeted nuclease technology. The complete GS knockout was confirmed via sequencing, glutamine sensitivity, and viability assays. CleanCut GS CHO cells demonstrate exceptional viability and recovery times, with doubling times of less than 20 hours, while achieving industry-leading cell specific productivity exceeding 70 pg/cell/day (Qp).
Key Features
- GS knockout achieved using the ultra-precise Cas-CLOVER technology — a clean alternative to CRISPR/Cas9.
- Industry-leading productivity with >70 pg/cell/day (Qp).
- Demonstrated capability with challenging-to-express monoclonal antibodies (mAbs), such as Rituximab.
- Robust growth parameters include viability, recovery, and rapid doubling times.
- The cell line history package demonstrates manufacturing processes consistent with regulatory compliance, featuring automated single-cell clonal visual reporting.
- Potential for integration with Demeetra gene editing IP and reagents for Cas-CLOVER or transposases for advanced cell line development.
Guide RNA strategy and characterization of mutants. The top section of the image shows the binding sites of a guide RNA pair within the CHO GS gene. Below this, sequence alignments are presented following Cas-CLOVER mediated targeting of GS in individual CHO clones. Significant deletions and frameshifts in exon 6, including 13, 22, and 25 base pair deletions, are depicted. Large deletions are commonly introduced using Cas-CLOVER, resulting in complete knockout phenotypes.
Sensitivity of CleanCut GS CHO clones to glutamine. Cell lines
show consistently decreased viability in medium without
glutamine, indicating a full knockout phenotype.
Monoclonality of CleanCut GS CHO clones. Single cell clones were derived using Solentim VIPS®.
Case Study: Trastuzumab (Herceptin)
Trastuzumab (Herceptin), a monoclonal antibody, is approved for the treatment of HER2-positive breast cancer. The heavy and light chain genes were stably integrated, with pools achieving an impressive >70 pg/cell/day (Qp). This successful demonstration underscores the potential of CleanCut GS CHO cells in streamlining cell line development.
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