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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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A New Dual Luciferase Assay Using NanoLuc® Enables a Second Generation Coincidence Reporter System to Reduce False Hits in HTS Poster
Christopher Eggers, Samuel Hasson, Brock Binkowski, Matt Robers, James Unch, Braeden Butler, , Keith Wood, James Inglese and Frank Fan

Luciferase-based reporter-gene assays remain a cornerstone of high-throughput screening of compounds because of their high sensitivity and dynamic range. However, a substantial number of non-relevant hits can be generated due to direct interaction of compounds with the luciferase reporter.

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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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iPSC-Derived Cardiomyocytes and Luciferase Reporters: A Robust Reporting Platform for Monitoring Cardioprotection and Pathway Biology in Endogenous Human Tissue Cells
Fiene, S., Thompson, A., Niles, A., Robers, M., Anson, B.

Pathophysiological conditions, medical interventions, and off-target toxicities can all result in cellular oxidative stress. In cardiac myocytes, prolonged and/or excessive oxidative stress can lead to cardiotoxicity: a primary cause of developmental delays, black-box warnings, and post-launch withdrawal of pharmaceuticals.

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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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The P450-Glo™ CYP2B6 Assay: a Rapid and Selective Assay for Measuring CYP2B6 Induction and Inhibition
Dongping Ma, Hui Wang, Poncho Meisenheimer, James J. Cali

We have developed a luminogenic CYP2B6 assay for biochemical CYP2B6 inhibition and for cell-based CYP2B6 induction studies. Here we present the CYP2B6 luminogenic assay characterization and demonstrate its utility for measuring time dependent CYP2B6 inhibition, and for measuring CYP2B6 induction in cultured primary human hepatocytes with normalization to viable cell count.

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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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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.
Genetic Mutation that Prevents Diabetes Complications
The most significant complications of diabetes include diabetic retinal disease, or retinopathy, and diabetic kidney disease, or nephropathy. Both involve damaged capillaries.
Charting Kidney Cancer Metabolism
Changes in cell metabolism are increasingly recognized as an important way tumors develop and progress, yet these changes are hard to measure and interpret. A new tool designed by MSK scientists allows users to identify metabolic changes in kidney cancer tumors that may one day be targets for therapy.
Individuals' Medical Histories Predicted by their Noncoding Genomes
Researchers have found that analyzing mutations in regions of the genome that control genes can predict medical conditions such as hypertension, narcolepsy and heart problems.
'Molecular Movie' Opens Door to New Cancer Treatments
An international team of scientists led by the University of Liverpool has produced a 'structural movie' revealing the step-by-step creation of an important naturally occurring chemical in the body that plays a role in some cancers.
Custom Tuning Knobs to Turn Off Any Gene
Factory managers can improve productivity by telling workers to speed up, slow down or stop doing tangential tasks while assembling widgets. Unfortunately for synthetic biologists attempting to produce pharmaceuticals, microbes don’t respond to direction like human personnel.
Unique Mechanism for a High-Risk Leukemia
Researchers uncovered the aberrant mechanism underlying a notoriously treatment-resistant acute lymphoblastic leukemia subtype; findings offer lessons for understanding all cancers.
Genetically Mapping the Most Lethal E.Coli Strains
New approach could lead to fewer deaths, and new treatments.
The Spice of Life
Scientists discover important genetic source of human diversity.
Cytoskeleton Crew
Findings confirm sugar's role in helping cancers survive by changing cellular architecture.
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