Articles, announcements and insights from Horizon Discovery

CRISPR technology raises standards for tracking protein expression

Apr 6, 2017 4:54:33 PM No Comments

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.

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Top peer reviewed scientific articles using HAP1 cell lines

Mar 13, 2017 10:24:39 AM No Comments

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. 

2017

2016

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Probing DNA Damage Response Pathways

Mar 8, 2017 1:02:47 PM No Comments

Sensitivity and resistance to DNA Damage Response Pathways identified with gene-edited cell lines and wildtype controls

The cellular DNA damage response (DDR) is an essential safeguard against cancer. Upon activation, the DDR can limit tumour progression at the early stages by inducing senescence or cell death. When this defence fails tumors are able to develop. However, with time, tumors accumulate more mutations in DNA repair proteins as cancers progress.  The efficiency of DDR plays an essential role in the effectivity of cytotoxic treatments. Currently much research is focussed on identifying the DDR mechanisms involved in cancers and how these dysfunctional processes can be utilized against tumor growth.

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How to Cure Cancer in 3 Easy Steps

Mar 6, 2017 6:10:15 PM No Comments

When I have read articles just like this one early on in my career, I would laugh and categorize it with blogs regarding Bigfoot and the Loch Ness Monster. However, much has changed in the past 10 years. New technologies have been developed and milestones have been reached that should have Cancer a little worried. These 3 steps might be viewed to some as obvious, but I argue that it’s how the researchers have utilized the technology wisely that has made the difference. I have identified some papers that have carved a successful path to Cancer's possible demise.

Read More Cell lines, Gene editing, CRISPR, Oncology Panels, Target identification, Target validation, PDX, Cancer models, Preclinical efficay testing, High Fidelity Models

Spying on proteins - How observation can affect the perception of reality

Mar 2, 2017 2:47:38 PM No Comments

One of the highest ideals in science is to observe natural events in their native context. Doing so is a constant challenge thanks to the Observer Effect, described by Heisenberg and others, where the act of observing or measuring a process alters it. Thus, scientists of all stripes try to get out of the way, attempting to produce the most accurate measurements possible using specific yet unobtrusive tools. Wildlife photographers use long-range lenses to avoid the need to stand directly in front of a herd of water buffalo and thereby affect the animals’ behaviour. Psychologists create tests where the subjects are unaware of the true intent so as to minimize changes in natural responses.

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Large scale gene tagging simplified by a self releasing plasmid

Mar 2, 2017 2:19:09 PM No Comments

Knockout and tagged gene-editing can create cell lines ideal for antibody validation. Using CRISPR CAS technology, Horizon developed a streamlined process to cut production time. Read more in our interview with Lead Scientist Daniel Lackner

Audio also available

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Endogenously tagged reporter cell lines to improve antibody validation

Mar 1, 2017 8:29:23 PM No Comments

CRISPR gene editing technology is being used to solve issues concerning how to localize intracellular proteins. Learn more by reading our interview with expert in this field,  Dr. Emma Lundberg

Audio also avaliable

David Shifrin: Welcome to Horizon Discovery. I'm David Shifrin. Today I'm speaking with Dr. Emma Lundberg, Associate Professor at the KTH Royal Institute of Technology and the Science for Life Laboratory. Doctor Lundberg, as we'll discuss in this conversation has a deep interest in looking at protein localization and expression throughout the many tissues of the human body during health and disease, and this comes out in the context of different cell lines in her research. Because of that interest, she's been involved in a number of different projects to ensure that the tools available to researchers for labeling proteins are really well-validated.

Then on the other side of things, Horizon Discovery is building a suite of endogenous pathway tag and reporter cell lines, the idea being that researchers can look at natural levels of protein and promoter activity with fewer artifacts than you'd expect from something like a traditional transfection reagent. The goal is really to help with projects like those that Dr. Lundberg is spearheading in the drive for quality in the antibody arena as well as a number of other projects that researchers might conceive.

With that, Dr. Lundberg, thanks very much for taking the time to speak with me today. How are you?

Emma Lundberg: I'm doing very well, thank you.

Mapping the localization of all human proteins

David Shifrin: Great. I'd like to start kind of big picture with your interest in this whole realm of antibody validation. You were part of a science paper published in early 2015 that mapped protein expression across more than 30 human tissues, and then late 2016 your lab published a paper in the Journal of Proteome Research that was a really interesting paper using endogenously expressed, tagged proteins to validate antibodies that were available through the Human Protein Atlas. Clearly this idea that ensuring that all the tools that researchers use to look at protein expression are very well validated, as I said before, and appropriate for the experiment at hand. Can you tell us a bit more about your interest in this idea and then kind of where all of that came from?

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Patient Derived Xenografts:  The Crucial Piece for Curing Cancer

Jan 4, 2017 3:57:00 PM No Comments
Read More PDX, preclinical efficacy testing, Cancer models, High Fidelity Models

Scientific Reports: Increasing the performance of pooled CRISPR–Cas9 drop-out screening

Nov 18, 2016 10:44:56 AM No Comments

Recently published in a paper on Nature.com in Scientific Reports, Horizon Discovery have conducted a detailed analysis of CRISPR-Cas9 sensitivity (drop-out) screening to come up with a highly improved and optimised platform. In our analysis, we used a custom ultra-complex sgRNA library and capitalised on Horizon’s streamlined screening pipeline to evaluate fundamental aspects of functional genomic screening, including:

  • Side-by-side comparison of the impact of a novel tracrRNA sequence on screen performance
  • Direct analysis of the efficacy of two different sgRNA design algorithms
  • Evaluation of the effect of cell line ploidy on KO rate and screen quality
  • Time-resolved gene drop-out analysis to evaluate the kinetics of CRISPR-Cas9 driven gene knockout
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Is there a better way to visualize proteins in cells?

Nov 17, 2016 1:37:26 PM No Comments

The visualisation of proteins or organelles in cells and other complex biological systems is ‘bread and butter’ stuff for cellular and molecular biologists; it’s performed day-in, day-out in labs across the globe. But this doesn’t mean that the most popular approaches currently used for protein visualisation – dye staining, antibody labelling and fusion protein over-expression systems – are ideal. Almost every scientific technique has its advantages and drawbacks, and the various options currently available for protein visualisation are not exceptions to this rule, so let’s have a look at the pros and cons of each.

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Spying on proteins - How observation can affect the perception of reality

One of the highest ideals in science is to observe natural events in their native context. Doing so...