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 HeLa cells can be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines.
Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
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Synonyms
SpCas9 cell pool, CRISPR cell pool, Cas9 cell pool, HeLa cell pool
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Product Data Gallery
Flow cytometry analysis of intracellular expression of Cas9 in HeLa cell pool
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.
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.
Thaw Medium 1 (BPS Bioscience #60187): DMEM medium supplemented with 10% FBS, and 1% Penicillin/Streptomycin
Growth Medium 1N (BPS Bioscience # 79801): DMEM medium supplemented with 10% FBS, and 1% Penicillin/Streptomycin plus 0.25 μg/mL Puromycin to ensure recombinant expression.
Cells should be grown at 37°C with 5% CO2 using Growth Medium 1N to ensure recombinant expression is maintained.
Instructions for Use
See assay protocol for detailed instructions.
Applications
1. Quickly generating knock-out cell pools or cell lines in HeLa cells. 2. Implementing sgRNA screens in Cas9-expressing HeLa cells
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.
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