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Application: 1. Quickly generating knock-out cell pools or cell lines in MDA-MB-231 cell pools.
2. Implementing sgRNA screens in Cas9 expressing MDA-MB-231 cell pools.
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 MDA-MB-231 cells can be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines.
Description: This cell line is a clonal derivative from the Cas9-Expressing MDA-MB-231 Cell Pool (BPS Bioscience, #78069). It was generated by limited limiting dilution of the original pool and isolation of individual clones, which were screened based on Cas9 expression to obtain a low- or high-expressing cell line. The expressed Cas9 protein includes a C-terminal FLAG tag.
Host Cell Line: MDA-MB-231
Mycoplasma Testing: This cell pool 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.
Supplied As: Each vial contains ~2 x 10^6 cells in 1 ml of FBS with 10% DMSO.
Biosafety Level: BSL-1
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