Articles, announcements and insights from Horizon Discovery

CRISPR-Cas9 is a very versatile tool to discover more about your pathway or gene of interest. After all that hard work editing your cell line, you want to have the confidence to rely on your new research model. So, how do you verify your cell line is what you expect it to be?

There are a couple of areas to consider:

• Cell line clonality - a mixed population may obscure the effects of your desired gene edits
• Is the targeted gene edit affecting the cell line phenotype, or is this due to off-target effects?

Here are a few ways to add supporting data to validate your gene-engineering projects...

CRISPR-edited Cell Lines are a great tool to validate your antibodies before you start your experiments. They ensure you are using high quality reagents, so you can be confident in your results. We explain how you can use Cell Lines for validation, the challenges to be aware of and how we can help you overcome them.

A major study has been undertaken to gain a better understanding of thousands of mutations in the BRCA1 gene – a key gene in breast and ovarian cancers. 

Published in Nature this month, the study by the Department of Genome Sciences at the University of Washington School of Medicine, set out to analyze nearly 4,000 variants  in 13 exons of BRCA1 that are “of unknown significance”. These are variants that are not currently known to cause cancer, but theoretically could. 

With over 25,000 individual Cell Lines in our new Express Catalog, we want to give you a helping hand with finding the ideal Cell Line Model for your research. 

Using our new downloadable catalog, you can now search, filter and browse through our online cell model offering. There are two main collections available in our catalog: 

• Knockout Cell Models
• Cancer-relevant Cell Lines

To help you find the right cell line for your research, here's some bite-size information on the data available to you... 

CRISPR technology now allows genes and molecular pathways to be examined with greater definition. We look at how knockout cell lines, either together with gene rescue and replication of disease mutations or as an independent cell model, can be used to validate your research and extend your findings.

There is a significant challenge in translating the wealth of genetic information now available to the role of genes, to understand basic biology, as well as linking to the role of mutations for understanding disease pathogenesis.

Knockout (KO) cell lines are excellent model systems to do this. A key benefit of cell lines is the ability to use gene-editing to construct isogenic cell line pairs - where a mutant model can be interrogated alongside a wildtype control.

Here’s five great examples of how to get the most out of your research using KO Cell Lines:

1. Validate your research tools
2. Identify key players in a pathway
3. Confirm pharmacological effects
4. Lift the quality of your research with proper controls
5. Connect patient relevant mutations to pathology

For the first time, Human Knockout cell lines are readily available for scalable reverse genetic screening.

We speak to Horizon's Head of Innovation, Dr Tilmann Bürckstümmer about the application of  reverse genetic screening using a combination of new technologies.

Gapp et al. (2016) in Molecular Systems Biology

Revealing the role of E3 ubiquitin ligases in DNA damage repair

One of the diverse new uses for the HAP1 cell line, one that has begun to draw significant attention, is in the field of DNA damage repair. A recent paper from Minoru Takata’s group highlights this important application of this relatively new tool.

The dilemma of when to invest in new technology

Researchers in the life sciences community are constantly walking a fine line in assay development. On one side is the accuracy, specificity and reproducibility borne from use of a well-established tool; i.e., a tool that has been on the market for a long time. Put another way, there is a level of comfort in using the same products for many years - in science as in the rest of life.

On the other side is the importance of finding the most efficient, cost-effective methods to carry out experiments. Doing so often means taking a chance on a new product, running it alongside existing methods to compare. Of course, it’s not just cost-effectiveness that necessitates making changes; simply keeping up can mean bringing in a new product that incorporates new advances. The outcome, hopefully, is better results faster, at lower cost.

And yet, inertia is a challenging force to overcome, and there is always a tendency to maintain the status quo. Particularly, as noted above, when so much rides on maintaining consistent protocols.

Here at Horizon, our scientists have built a remarkable new tool in the HAP1 cell line to facilitate researcher's access to CRISPR technology. These knockout cell lines allow researchers to quickly validate their gene or target of interest, without having to invest time and resource in developing in-house CRISPR technology.

HAP1 and HAP1 cells gene-edited to knockout SLC30A6 (HAP1_SLC30A6, catalogue number: HZGHC002784c010) with the HPA antibody HPA057328 targeting SLC30A6 demonstarting the specificity of this antibody. The samples were prepared in parallel using the same antibody dilutions and reagents, and both images are acquired with the exact same settings. Images curtesy of Dr Emma Lundberg, Cell Profiling facility. KTH Royal Institute of Technology.

We believe that, for its designated applications, HAP1 cells are more than worth adding into a lab’s toolbox. However, our opinion only takes things so far. So we set out to ask a few scientists who have published using the HAP1 cells about their work, and how the cells played a role in their investigations.

Read our blog on how new methods increase precision in protein visualization

Here we describe some of the great solutions that are coming out of recent advances using CRISPR CAS technology that give more precise and physiological results for protein visualization.  In our previous blog (see link at end of article), we discussed the some of the difficulties  with traditional methods for protein tracking and localization. One of the main causes of variability and wasted resources is the lack of standards for antibody quality. 

To be useful, an antibody must:

• Be specific

• Have a high signal to noise ratio

• Be validated for the assay at hand

Efforts to reduced non-specific antibodies in both industry and academia

From an industry standpoint, numerous organizations and commercial suppliers have created (or are creating) programs to ensure that the above criteria are met for each new antibody brought to market.

Can I use the HAP1 cell line for my research?

The HAP1 cell line has been applied across a wide range of biological processes, such as DNA damage repair pathway and stress responses, as well as in disease modeling. These selected articles show the broad applicability of the HAP1 cell line, and provide characterization data to help your research. If you would like to know more, please follow these links to our ready made cell lines and cell line engineering services.