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Picking the best CRISPR-Cas9 targets for functional gene knockout: a machine learning algorithm based on both specificity and functionality
Shawn McClelland, Emily M. Anderson, Žaklina Strezoska, Elena Maksimova, Annaleen Vermeulen, Steve Lenger, Tyler Reed, and Anja van Brabant Smith Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US

The CRISPR-Cas9 system has the potential to significantly advance basic and applied research.

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Predicting Regioselectivityand Labilityof Cytochrome P450 Metabolism using Quantum Mechanical Simulations
Tyzack, Nicholas Foster, Peter Hunt, Matthew Segall

Predicting Regioselectivity and Lability of Cytochrome P450 Metabolism using Quantum Mechanical Simulations

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Scaffold design, function and over-expression of lentiviral-based microRNAs
Angela Schoolmeesters, Melissa L. Kelley, Annaleen Vermeulen, Anja Smith, *Mayya Shveygert, *Xin Zhou, *Robert Blelloch Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, USA

Here we describe the strategy for scaffold design, the importance of an optimal promoter, and demonstrate gene target down-regulation from the over-expression of lentiviral microRNA mimics.

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Homology-directed repair with Dharmacon™ Edit-R™ CRISPR-Cas9 and single-stranded DNA oligos
John A. Schiel, Eldon T. Chou, Maren Mayer, Emily M. Anderson , and Anja van Brabant Smith | Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US

Here we demonstrate how to perform lipid based transfections for homology directed repair using DharmaFECT Duo, CRISPR-Cas9 reagents and, synthetic DNA donor oligos.

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Minimizing Carry-over for High Throughput Analysis
Christian Berchtold1, Reto Bolliger2, Guenter Boehm2, Götz Schlotterbeck1

Minimal carry-over is a prerequisite for high throughput analysis. However, minimized carry-over and cycle time are competing and a careful optimization is mandatory. In this study the influence of wash conditions on carry-over of various compounds was investigated. A strategy to minimize carry-over was developed. The influences of different wash tasks were investigated. Finally the contribution of different system components such as injector valve or column was studied.

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A Comparison of ITEX Dynamic Headspace–GC/MS to other Enrichment Techniques for Analysis of Flavoring Compounds
Douglas Doster1; Roger Pearson1; Sean Eppel1; Ken Rice1; Tom Flug2; Brian Peat2; Guenter Boehm2

Enrichment techniques are commonly used for the analysis of flavoring compounds in different matrices with GC/MS. Analysis of flavoring compounds is done by purge & trap, SPME or headspace, depending on requirements for sensitivity. The In-Tube Ex¬traction (ITEX) Dynamic Headspace uses a micro trap filled with an adsorbent material to efficiently extract the compounds. Here we evaluate if the ITEX can be used to effectively analyze for these compounds and reduce the analyst’s time involved.

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Automated in-gel digestion on a commercial autosampler directly coupled to nanoLC-MS/MS
Achermann François, Bolliger Reto, Buchs Natasha, Doiron Nicholas, Lagache Braga Sophie, Heller Manfred, Boehm Guenter

SDS-PAGE separates protein samples from LC-MS incompatible contaminations, and is frequently used to fractionate proteins of entire proteomes. One disadvantage is that gel lanes have to be cut into many slices, followed by in-gel digestion of proteins and extraction of peptides. The number of these gel slices goes into the hundreds, rendering this process very repetitive and prone to mistakes and errors during sample handling. Automation reduces such risks and improves reproducibility.

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Automated sample preparation workflows for quantitative proteomics applications
Oliver Popp1, Lucas Luethy2, Tamara Kanashova1, HaAn Nguyen1, Julia Kikuchi1, Guenter Boehm2, Thomas Blenkers3, Andreas Bruchmann3, Gunnar Dittmar1

Mass spectrometry based proteomics requires large scale identification of peptides, and depends upon efficient sample preparation. Recently, we presented two automated protein-digestion setups, in-solution and in-gel digestion. We extended these techniques by implementing dimethyl labelling (DML). Furthermore, we established an automated phospho-peptide (PP) enrichment procedure in a 96-well formate, generating phospho-proteomic data in very short time.

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Tools for studying and using small RNAs: from pathways to functions to therapies
Kenneth Chang and Gregory J. Hannon

This poster provides an overview of the tools that have been developed to understand the functions of small RNAs and, conversely, the use of small RNAs as tools. Tools that are based on small RNAs have been exploited to investigate gene function in cultured cells and in living animals. Small RNA biogenesis, discovery and functional roles are explored in detail. Screening approaches to functional genomics, in vivo methods and potential therapeutic applications are discussed.

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Scientific News
Flowering Regulation Mechanism Discovered
Monash researchers have discovered a new mechanism that enables plants to regulate their flowering in response to raised temperatures.
Turning Skin Cells into Heart, Brain Cells
In a major breakthrough, scientists at the Gladstone Institutes transformed skin cells into heart cells and brain cells using a combination of chemicals.
Nanoparticles Present Sustainable Way to Grow Food Crops
Nanoparticle technology can help reduce the need for fertilizer, creating a more sustainable way to grow crops such as mung beans.
How Scientists Use DNA to Track Disease Outbreaks
They’re the top questions on everyone’s mind when a new disease outbreak happens: where did the virus come from? When did this happen? How long has it been spreading in a particular country or group of people?
Genetic Risk Factors of Disparate Diseases Share Similar Biological Underpinnings
Penn Institute for Biomedical Informatics and colleagues identify "roadmap" of disease mechanisms to identify candidate drug targets.
Drugs that May Combat Deadly Antibiotic-Resistant Bacteria Uncovered
Study identifies 79 compounds that inhibit carbapenem-resistant Enterobacteriaceae (CRE).
Stem Cells Know How to Unwind
Research led by the Babraham Institute with collaborators in the UK, Canada and Japan has revealed a new understanding of how an open genome structure supports the long-term and unrestricted developmental potential in embryonic stem cells.
HIV Particles Used to Trap Intact Mammalian Protein Complexes
Belgian scientists from VIB and UGent developed Virotrap, a viral particle sorting approach for purifying protein complexes under native conditions.
Childhood Asthma Research Receives $2M
Research into the impact of a child’s upbringing and social and physical environments on the development of asthma will receive $2 million to tackle the condition that affects as many as one in three Canadians.
Growing Stem Cells More Safely
Nurturing stem cells atop a bed of mouse cells works well, but is a non-starter for transplants to patients – Brown University scientists are developing a synthetic bed instead.
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