|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.
|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.
|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.
|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.
|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.
|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.
|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.
|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.
|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.