CHOZN® cells (GS−/− CHOK1, Sigma) were passaged regularly in EX‐Cell CD CHO Fusion medium (Sigma 14365C) containing 6 mM Glutamine. Per gRNA transfection, 1ug TrueCut Cas9 V2 proteins (Thermo Fisher A36496), 200 ng CRISPR IVT gRNA (Table 1 ), and 10 pmol ss‐Oligos (Table 1 ) in 10ul reaction volume were electroporated into 3.0E + 6 cells using NEON electroporator, following the optimized parameters of 1,600 V pulse voltage, 10 ms each pulse for three pulses. In 72 hr posttransfection, a portion of transfection cells were processed for the estimation of gene editing efficiency using GeneArt genomic cleavage detection kit (Thermo Fisher A24372). The rest of cells were cultured to expand for stable pool generation. The pools of highest editing efficiency were single cell sorted into 10 plates of 96‐well plates. Once confluent, the clones were consolidated to one 96‐well plate and gRNA‐targeted genomic regions were polymerase chain reaction (PCR) amplified (Table 1 ) and sequenced with next generation sequencing.
Chozn cells
Manufactured by Merck Group
The CHOZN® cells are a proprietary Chinese Hamster Ovary (CHO) cell line developed by the Merck Group. They are designed for use in biopharmaceutical production and research applications. The CHOZN® cells provide a stable and high-yielding system for the expression of recombinant proteins.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using chozn cells
1
CRISPR-Cas9 Mediated Gene Editing in CHOZN Cells
2
CHOZN Cell Galactose Induction
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In a separate experiment, the CHOZN cells (Sigma–Aldrich; St. Louis, MO) were grown in the wells of deep 12‐well plates. Like the perfusion reactor the cells were grown in a 90/10 blend of the Platform Media/Feed. The cells were treated with either 1, 5, or 10 mM galactose. Following 4 days in culture the cells were harvested, centrifuged, and the pellet flash frozen at −80°C until Western blot analysis.
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