This cell line is a clonal derivative from the Cas9-Expressing Raji Cell Pool (BPS Bioscience, #78071). It was generated by limiting dilution of the original pool and isolation of individual clones, which were screened based on Cas9 expression to obtain a high-expressing cell line. The expressed Cas9 protein includes a C-terminal FLAG tag.
Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
This cell line has been screened using the MycoAlert™ Mycoplasma Detection Kit (Lonza, #LT07-118) to confirm the absence of Mycoplasma contamination. MycoAlert Assay Control Set (Lonza, #LT07-518) was used as a positive control.
Background
Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease enzyme that, when recruited to a specific DNA sequence by the sgRNA (single guide RNA), introduces a double stranded break into the DNA. This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the targeted gene. Cas9-expressing Daudi cells can be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines.
Storage/Stability
Cells will arrive upon dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.
Culture Conditions
For best results, it is highly recommended to use these validated and optimized media from BPS Bioscience. Other preparations or formulations of media may result in suboptimal performance.
Growth Medium 2K (BPS Bioscience, #78078): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin plus 0.25 μg/ml of Puromycin to ensure recombinant expression.
Cells should be grown at 37°C with 5% CO2 using Growth Medium 2K to ensure recombinant expression is maintained.
Troubleshooting Guide For all questions, please email [email protected].
The CRISPR/CAS9 technology is covered under numerous patents, including U.S. Patent Nos. 8,697,359 and 8,771,945, as well as corresponding foreign patents applications, and patent rights.