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Design considerations for highly specific and efficient synthetic crRNA molecules
Anja van Brabant Smith, Emily M. Anderson, Shawn McClelland, Elena Maksimova, Tyler Reed, Steve Lenger, Žaklina Strezoska, Hidevaldo Machado Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US

An overview of our rational design algorithm for picking highly functional crRNA sequences in combination with comprehensive specificity analysis.

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CellTiter-Glo® 2.0: A Novel Luminescent Cell Viability Assay with Greatly Enhanced Storage Stability
Michael P. Valley, James Unch, Poncho L. Meisenheimer, James J. Cali, and Dan F. Lazar

Here we report on the attributes of a novel ATP detection reagent for cell viability with all of the assay performance of the previous CellTiter-Glo® Reagent, but now with markedly enhanced stability as a single component in a liquid format. These new features provide for much greater ease-of-use in that storage of the reagent at 4°C eliminates the requirement for reagent thawing and minimizes temperature equilibration time.

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Design and Validation of Bioluminescent Assays for 3D Cell Culture Models Poster
Terry L. Riss, Michael P. Valley, Chad A. Zimprich, Andrew L. Niles, Kevin R. Kupcho and Dan F. Lazar

Cells cultured in 3D model systems often acquire relatively large in vivo-like structures compared to the thickness of a 2D monolayer of cells grown on standard plastic plates.

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Testing a Novel Real Time Cell Viability Assay
Amy Landreman, Sarah Duellman, Wenhui Zhou, Jolanta Vidugiriene, Brad Hook

Recently developed assay technologies make it possible to use multi-well plate readers to measure the number of live or dead cells in culture in real time over a period of days. Live cells are measured in real time by adding a reagent containing a shrimp-derived luciferase and a pro-substrate directly to the culture medium. Only viable cells can convert the pro-substrate into a luciferase substrate and generate light.

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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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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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Increasing gene editing efficiencies in eukaryotic cell lines by selection of appropriate CRISPR-Cas9 reagents
Melissa L. Kelley, Žaklina Strezoska, Elena Maksimova, Hidevaldo Machado, Emily M. Anderson, Maren Mayer, Annaleen Vermeulen, Shawn McClelland, Anja van Brabant Smith

Overview of various CRISPR-Cas9 reagents to provide the highest efficiency of gene editing in your experiments.

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Knockdown of p53 by Accell self-delivering siRNA causes inhibition of p53-dependent DNA damage response in IMR-32 neuroblastoma cell line and β-amyloid toxicity in rat cortical neurons
Žaklina Strezoska, Tamara Seredenina1, Devin Leake, Annaleen Vermeulen

Here we describe how application of Accell siRNA enabled the development of a high content screening assay in IMR-32 neuroblastoma cells and a whole culture cell viability assay in primary rat cortical neurons.

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An Efficient Method for the Incorporation of Molecular Probes at Multiple/Specific sites in RNA: Levulinyl Protection for 2'-ACE ® , 5'-Silyl Oligoribonucleotide Synthesis
Xiaoqin Cheng, Shawn Begay, Randy Rauen, Kelly Grimsley, Kaizhang He, Michael Delaney

A unique method that uses a levulinate ester as a protecting group to introduce conjugates or molecular probes to virtually any location in a synthetic RNA molecule is discussed. The Levulinyl protecting group is stable in RNA synthesis conditions and can be removed without affecting the other parts of the synthesized RNA. We show the capabilities of this approach with three high-complexity synthesis examples.

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Scientific News
Common Cell Transformed into Master Heart Cell
By genetically reprogramming the most common type of cell in mammalian connective tissue, researchers at the University of Wisconsin—Madison have generated master heart cells — primitive progenitors that form the developing heart.
Mitochondria Shown to Trigger Cell Ageing
An international team of scientists has for the first time shown that mitochondria, the batteries of the cells, are essential for ageing.
Cancer Cells Kill Off Healthy Neighbours
Cancer cells create space to grow by killing off surrounding healthy cells, according to UK researchers working with fruit flies.
Editing of Embryos Approved in the UK
The Human Fertilisation and Embryology Authority (HFEA) has approved a research application from the Francis Crick Institute to use new "gene editing" techniques on human embryos.
Microbes Take Their Vitamins
Scientists exploit organisms' needs in order to track 'vitamin mimics' in bacteria.
Machine Learning Uncovers Unknown Bacterial Features
Technique robustly identified characteristic gene expression patterns in response to antibiotics, low oxygen conditions.
CRISPR-Cas9 Gene Editing Advances Again
UC Berkeley researchers have made a major improvement in CRISPR-Cas9 technology that achieves an unprecedented success rate of 60 percent when replacing a short stretch of DNA with another.
Disrupting Cell’s Supply Chain Freezes Cancer Virus
When the cancer-causing Epstein-Barr virus moves into a B-cell of the human immune system, it tricks the cell into rapidly making more copies of itself, each of which will carry the virus.
Why Do Some Infections Persist?
In preparing for the possibility of an antibiotic onslaught, some bacterial cultures adopt an all-for-one/one-for-all strategy that would make a socialist proud, University of Vermont researchers have found.
ASCB: A CELLebration of Cell Biology
The last major congress of the year, ASCB is less a platform for launching new products, but one for confirming and consolidating the trends that have emerged over the past 12 months.
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