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Pathfinder Target Essentiality Assay Service A new CRISPR─Cas9 based medium throughput assay service for validation of target gene essentiality Can be used to resolve ambiguous screening results Can also provide information on drug target suitability This assay developed at Horizon will enable you to identify genes essential for the growth of specific cancer cell lines. It can be used to definitively resolve ambiguous screening results. Or to provide information on target suitability – by testing essentiality in “normal” cells, or in cancer subtypes different to the proposed patient population

Resolving Ambiguity in Target ID Screens - CRISPR-Cas9 Based Essentiality Pro...

Candy Smellie

Manual assessment of biomarker expression is associated with significant inter- and intra reader variability. In some cases there are also limitations when it comes to sensitivity and specificity of manual biomarker assessment. In one example to the left, the “pure” contribution of inter-reader variability associated with Ki67 assessment was quantified across 20 tumors and 126 participating labs. In that study, it was demonstrated how image analysis can be used to significantly reduce inter-reader variability. In a another study, the National Danish Validation study of Her2, it was demonstrated how improved sensitivity/specificity of quantitative HER2 protein expression wrt gene amplification lead to significant cost savings in reflex testing. By automating aspects of stain quality control, it will become scalable to he point where EQA organizations may be able and willing to offer more frequent – perhaps even on-demand – proficiency testing and calibration services. It is possible that objective and quantitative standards will contribute to improve compliance with protocol recommendations. In clinical multi-center trials it will be easier to standardize and monitor data from each center. And it is our hope tha larger diagnostic pathology labs will be able to benefit from such a method by closely monitoring drift in staining quality for biomarkers.

Blueprints to blue sky – analyzing the challenges and solutions for IHC compa...

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CRISPR/Cas9 gene editing is based on a microbial restriction system, that has been harnessed for genome targeting using only a short sequence of RNA as a guide. The beauty of the system is that unlike protein binding based technologies such as Zinc Fingers and TALENs which require complex protein engineering, the design rules are very simple, and it is this fact that is allowing CRISPR to take genome engineering from a relatively niche persuit to the mainstream scientific community. The principle of the system is that a short guide RNA, homologous to the target site recruits a nuclease – Cas9 This then cuts the dsDNA, triggering repair by either the low fidelity NHEJ pathway, or by HDR in the presence of an exogenous donor sequence. High Efficiencies for both knockouts and knock-ins have been reported and whilst there are understandable concerns about specificity, new methodologies to address these are now being developed The system itself is comprised of three key components the Cas9 protein, which cuts/cleaves the DNA and Two RNAs - a crispr RNA contains the sequence homologous to the target site and a trans-activating crisprRNA (or TracrRNA) which recruits the nuclease/crispr complex For genome editing, the crisperRNA and TraceRNA are generally now constructed together into a single guideRNA or sgRNA Genome editing is elicited through hybridization of the sgRNA with its matching genomic sequence, and the recruitment of the Cas9, which cleaves at the target site.

CRISPR: Gene editing for everyone

Candy Smellie

Genome Editing Comes of Age

Candy Smellie

Information is no longer a bottleneck, emphasis is shifting to the ‘what does it all mean’ In a translational context we hope that by answering that question we will be able to is to characterise the genetics that drive disease, and indeed develop drugs and diagnostics that are personalised to patients. Genome editing provides the link between the information here, and this outcome here, by allowing scientists to recapitulate specific genetic alterations in any gene in any living tissue to probe function, develop disease models and identify therapeutic strategies. So, not only do we now have unparalleled access to genetic information, but we now have the tools to most accuartely understand what this genetic information – with genome editing allowing us to explore the genetic drivers of disease in physiological models. AAV is a single-stranded, linear DNA virus with a a 4.7 kb genome which for the purpose of genome editing is replaced almost in entirety with the targeting vector sequence (except for the iTRs) It is in effect a highly effective DNA delivery mechanism After entry of the vector into the cell, target-specific homologous DNA is believed to activate and recruit HR-dependent repair factors can induce HR at rates approximately 1,000 times greater than plasmid based double stranded DNA vectors, but the mechanism by which it achieves this is still largely unknown By including a selection cassette can select for cells that have integrated the targeting vector, and then screen for clones which have undergone targeted insetion rather than random integration, which will generally be around 1%.

Genome Editing Comes of Age; CRISPR, rAAV and the new landscape of molecular ...

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Resolving Ambiguity in Target ID Screens - CRISPR-Cas9 Based Essentiality Pro...

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Isadora Agency and the UPLI team wanted to redefine how employees especially millennials engaged with own finances. In short, business goals included helping users build credit, maintain a budget, optimize debt and manage all financial accounts. The final product was something truly unique. Simple enough to track finances much like you track your health. Web design specialists at Isadora Agency were tasked with the app UI design, mobile strategy, illustration, iconography, and mobile app design of over 100 different mobile screens With a minimalist design, users quickly move from one category to the next. For example, you can select to pay down your debt or fund a vacation. Therefore, your personal dashboard allows you to see insights, progress, and trends. The Budget Progress screen was extra fun to design as we were able to incorporate our custom character illustration next to messages of encouragement. It’s truly remarkable how the team packed this mobile app design with so many tools and powerful customizable features. The core of the app UI design revolves around essential features that include budgeting, spending, and saving. For example, layers of friendly reminders and updates show how much you’ve saved or how close you remained within your budget. Meanwhile, providing multiple reasons to visit the app often. For instance, cutting-edge tools and intelligent messaging throughout the experience educate users as they grow more comfortable with individual finances.

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