GMP GENPowerTM NLS-Cas9 Nuclease is a recombinant Streptococcus pyogenes Cas9 protein, purified from Escherichia coli for CRISPR-based genome editing. The addition of nuclear localization signals (NLS) enhances nuclear entry, increasing the likelihood of genomic DNA cleavage.
Introduction
Features
- Designed under ISO 9001:2015 and ISO 13485:2016 standards
- Manufactured and QC tested in a GMP-compliant facility
- FDA DMF filed
- Animal-free materials
- Beta-lactam-free formulation
- Consistent batch-to-batch quality
- Stringent quality control testing
- No animal-derived peptone or lactose used in the production process
Application
- Genetic modification of cells and gene therapy applications, including T cells and hematopoietic stem cells
- Highly specific pathogen detection
Concentration
10 mg/mL
Purity
>95 %, as SDS-PAGE analysis revealed.
>95 %, as SEC-HPLC measured.
Host cell protein
Protein tested by ELISA: ≤10 ng/mg.
Host cell DNA
≤1 ng/mg protein as determined by qPCR.
Sterility
According to CP<1101>, USP<71>, and Eur. Ph. 2.6.1, the membrane filtration method was used to conduct the sterility test.
Endotoxin
LAL method: less than 10 EU/mg.
Formulation
Supplied in a 0.2 μm filtered solution with pH 7.5, 20 mM Tris, 300 mM NaCl, 0.1 mM EDTA, and 1 mM TCEP.
Shipping
This product is supplied and shipped with dry ice.
Storage
Once stored, this product is stable at:
- -20 °C for five years in a sterile environment
- When the product is received, it MUST be stored at -20 °C or lower
ACRO quality management system
- QMS: ISO, GMP
- Quality Benefits
- The process of quality control
QC and assay protocol
SDS-PAGE
Coomassie Blue was used to stain the gel. According to Star Ribbon Pre-stained Protein Marker, the protein's purity is higher than 95 %.
Image Credit: ACROBiosystems Inc
SEC-HPLC
According to SEC-HPLC, the purity of GMP GENPowerTM NLS-Cas9 Nuclease (Cat. No. GMP-CA9S18) was higher than 95 %.
Image Credit: ACROBiosystems Inc
Bioactivity
Different amounts of Cas9 were incubated with a fixed excess of gRNA and plasmid for 60 minutes at 37 °C. When using 400–200 ng Acro Cas9, the cutting efficiency exceeded 90 % (QC tested). In contrast, 200 ng of Competitor T achieved only around 50 % cutting efficiency.
Image Credit: ACROBiosystems Inc
In human primary T cells, GMP GENPowerTM NLS-Cas9 Nuclease's TCR knockout efficiency was over 95 %.
Image Credit: ACROBiosystems Inc
The cleavage efficiency in HEK293 cells 72 hours after electroporation of GMP GENPowerTM NLS-Cas9 Nuclease RNP.
Image Credit: ACROBiosystems Inc
The cleavage efficiency in iPSC 72 hours following GMP GENPowerTM NLS-Cas9 Nuclease RNP electroporation.
Image Credit: ACROBiosystems Inc
Using flow cytometry, the B2M knockout efficiency in primary T cells was determined.
Image Credit: ACROBiosystems Inc
Using flow cytometry, the TRAC knockout efficiency in Jurkat cells was determined.
Image Credit: ACROBiosystems Inc
Using flow cytometry, the B2M knockout efficiency in Jurkat cells was determined.
Image Credit: ACROBiosystems Inc
Bioactivity-Stability
Batch-to-batch consistency between PG Cas9 and ACRO's GMP is demonstrated by the bioactivity-based assay.
Image Credit: ACROBiosystems Inc