Knock-out cell lines

Understand disease mechanisms and validate potential therapeutic targets with knock-out (KO) cell lines

​Find your KO cell line

Knock-out (KO) cell lines help overcome the challenge of accessing the correct controls during assay development and enable confident interrogation of the relationship between genotype and phenotype. 

However, generating CRISPR KO cell lines is time-consuming, with an average of three to four repeats of the CRISPR workflow before they obtain the specific CRISPR edits required ¹. 

To give you this time back, we have established a large and expanding range of ready-made and made-to-order knock-outs in commonly used cell lines that are ready to go, making CRISPR knock-out cell lines as easy to buy as reagents. 

Our CRISPR knock-out cell line catalog is designed to cut more than four months from your preparation time, taking you from months of experimental work to a simple click to achieve the CRISPR knock-out cell line you need – meaning you can get more data and publish faster.

It's the quickest and most cost-effective option to accelerate your program.

• Benefits of our gene-edited cell lines
• How do we generate our gene-edited cell lines?
• FAQs

Benefits of our gene-edited cell lines

• Quickly find your target with our easy-to-navigate catalog and ordering process.
• Physiologically relevant and certified mycoplasm-free cell backgrounds.
• Controls are provided with all orders to help you produce robust, reproducible results.
• Genomically validated, and in many cases, proteomically validated to confirm full gene ablation at the genomic and proteomic levels. All this data is available with each specific CRISPR knock-out cell line.
• Our catalog covers over 1000 key targets in multiple backgrounds, including physiologically relevant backgrounds such as A549 and MCF-7.
• Validating antibodies using knock-outs is considered the gold standard for antibody validation.
• KO cell lines are powerful tools to study disease mechanisms and investigate the relationship between genotype and phenotype, making them suitable for identifying novel therapeutic targets.
• The CRISPR revolution has fueled a rapid expansion of gene-edited cells in a variety of assays, streamlining drug discovery and assay development.

How we generate our  gene-edited cell lines

1. Cell line selection

Every knock-out cell line starts with the selection of the most appropriate target gene and background cell line. Our selections are made in close collaboration with the research community to ensure that we’re always supporting our customers with the cell lines that are most relevant to their research and application needs. 

We obtain our background cell lines from standard repositories, such as the American Type Culture Collection (ATTC), which are fully validated and certified mycoplasma-free.

If we haven’t used a line before, we conduct a thorough evaluation to check suitability, including confirming that the protein of interest is expressed in the wild-type cells and ensuring that the cells will grow and remain viable after the target is knocked out.

In addition to using workhorse cell lines such as HEK and HeLa, we produce knock-outs in various physiologically relevant and disease-relevant backgrounds. For example, CD74 is highly expressed in lymphoma, so we generate our knock-out line for human CD74 in a Raji lymphoma cell line.

2. CRISPR guide design

Our high-throughput platform allows us to automate the production of our CRISPR knock-out cell lines at scale. This reduces the time typically required to produce gene-edited monoclonal cell lines, so you can gain back valuable lab time, have confidence in scalable and long-term supply, and achieve consistent results over time.  

Creating a successful CRISPR knock-out cell line depends on selecting suitable guide RNAs; our CRISPR guide RNAs are carefully designed and optimized using comprehensive web-based gRNA design tools – including alignment-based scoring and hypothesis-driven methods – to increase the accuracy and efficiency of targeting.

Crucially, we employ a dual RNA system, using two guides to recognize flanking sites around the target sequence rather than a single site. This generates a small fragment deletion, guaranteeing a complete protein knock-out and increasing our typical knock-out efficiency to over 95% compared with a single guide efficiency of around 55%. This reduces our screening time ^2,3 and translates into cost efficiency and reliability for our customers. 

3. Transfection

Once we’ve designed our RNA guides, the CRISPR components need to be transfected into cells to begin the editing process. 

Conventionally, the components of the CRISPR system are delivered into cells using DNA plasmids, relying on the cell to express the Cas enzyme and guide RNAs needed for editing. But this technique can introduce uncertainty because it relies on efficient transcription of all the CRISPR system components by the cell. It also allows the CRISPR system to be expressed continuously, increasing the potential for off-target edits, and leaves external plasmid DNA in the cells, which may cause problems down the line. 

We take a different approach. By delivering pre-assembled CRISPR ribonucleoprotein complexes directly into cells, we benefit from a transient editing system that dissipates, so there is no continuous CRISPR expression, and no extra genetic material left behind. As well as enhancing accuracy, this system allows us to chemically modify guide RNAs before transfection to increase specificity even further⁴.

 Once the editing process is complete, we use a clonal selection workflow (followed by an extensive validation process) to create a homologous knock-out cell line.

We perform CRISPR on bulk transfected cell ‘pools,’ which are then screened using next-generation and Sanger sequencing to confirm editing efficiency. Pools containing successfully edited cells are then single-cell-cloned and expanded to create clonal cell populations.

Frequently asked questions (FAQs)