Cas9-Expressing HCT116 Cell Line (High or Low Expression)

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Catalog #

78135

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Description

This cell line is a clonal derivative from the Cas9-Expressing HCT116 Cell Pool (BPS Bioscience, #78073). 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 (78135-H) or low-expressing (78135-L) cell line. The expressed Cas9 protein includes a C-terminal FLAG tag.

This product has been cited 1 time.

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 line, CRISPR cell line, Cas9 cell line, HCT116 cells, Human Colorectal Carcinoma Cells

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Product Data Gallery

Flow cytometry analysis of intracellular expression of Cas9 in HCT116 cells

Product Info

Storage and Usage

Citations1

Host Cell Line

HCT-116 human colorectal carcinoma cell line. Adherent epithelial cells.

Species

S. pyogenes

Supplied As

2 vials of frozen cells. Each vial contains 2 x 10⁶ cells in 1 ml of 10% DMSO

Materials Required But Not Supplied

Materials Required for Cell Line Culture
Name	Ordering Information
Thaw Medium 7	BPS Bioscience #60185
Growth Medium 7C	BPS Bioscience #78076

UniProt #

Q99ZW2

Mycoplasma Testing

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 HCT116 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.

Thaw Medium 7 (BPS Bioscience #60185): McCoy’s 5A medium with 10% FBS, 1% Penicillin/Streptomycin

Growth Medium 7C (BPS Bioscience #78076): McCoy’s 5A medium with 10% FBS, 1% Penicillin/Streptomycin plus 1 μg/ml Puromycin to ensure recombinant expression.

Cells should be grown at 37°C with 5% CO2 using Growth Medium 7C to ensure recombinant expression is maintained.

Instructions for Use

See assay protocol for detailed instructions.

Shipping Temperature

-80°C

Notes

License Disclosure
Visit bpsbioscience.com/license for the label license and other key information about this product.

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.

Warnings

Avoid freeze/thaw cycles.

Scientific Category

CRISPR

Regulatory Status

Research Use Only

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Cas9-Expressing HCT116 Cell Line (High or Low Expression)

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