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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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Innovative technology that enables RNAi in difficult to transfect cells
Christina Yamada, Kathryn Robinson, Allison St. Amand, Zaklina Strezoska, Greg Wardle, Anastasia Khvorova, Devin Leake

Investigations at Dharmacon have led to the development of innovative siRNA molecules that can be delivered into difficult-to-transfect cells without additional lipid reagents, virus, or instruments. This technology, Accell siRNA reagents, enables gene knockdown for functional genomic studies in a wide variety of cell types. In some instances, cells can be continuously dosed with Accell siRNAs to enable target gene knockdown for extended durations.

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Innovative technology that enables RNAi in difficult to transfect cells
Christina Yamada, Kathryn Robinson, Allison St. Amand, Zaklina Strezoska, Greg Wardle, Anastasia Khvorova, Devin Leake

Investigations at Dharmacon have led to the development of innovative siRNA molecules that can be delivered into difficult-to-transfect cells without additional lipid reagents, virus, or instruments. This technology, Accell siRNA reagents, enables gene knockdown for functional genomic studies in a wide variety of cell types. In some instances, cells can be continuously dosed with Accell siRNAs to enable target gene knockdown for extended durations.

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Modelling CLL cell and T-cell Migration in a Dynamic Circulating Model of CLL
Elisabeth Walsbyl, Paul Brennan', Guy Pratte, Andrea Buggins3, Tanja N Hartmann'', Chris Fegan1 and Chris Pepper'

We have recently developed a novel circulating model of chronic lymphocytic leukaemia (CLL) that mimics the transient interactions that take place between circulating lymphocytes and vascular endothelium. Here we show that both normal and malignant lymphocytes actively underwent transendothelial migration.

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Measuring Antitumor Effect of c-Myr-Max heterodimerization inhibitor 100258-F4 on Ovarian Cancer Cells using Cellometer Imaging Cytometry
Leo L. Chan, Jiandong Wang, Xiaoli Ma, Hannah M. Jones, Fang Song, Weiyuan Zhang, Victoria L. Bae-Jump, Chunxiao Zhou

The effect from the c-Mycinhibitor 100258-F4 was clearly observed from the G0/G1 cell cycle arrest and the increase in early and late apoptotic cell. The Cellometer method can measure fluorescent cell-based assays such as cell cycle, apoptosis, protein expression, autophagy and viability. The ability to export the data to FCS Express facilitates simple data analysis and reporting to generate results for publications.

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Scientific News
Shape Of Tumor May Affect Whether Cells Can Metastasize
Illinois researchers found that the shape of a tumor may play a role in how cancer cells become primed to spread.
‘Mini-Brains’ to Study Zika
Novel tool expected to speed research on brain and drug development.
Cytokine Triggers Immune Response at Expense of Blood Renewal
Research highlights promise of Anti-IL-1 drugs to treat chronic inflammatory disease.
Micro Heart Muscle Created from Stem Cells
Researchers have designed a new way to create micro heart muscle from stem cells using a unique dog bone dish.
“Secret Sauce” for Personalized, Functional Insulin-producing Cells
Researchers uncover molecular switch to make effective sugar-responsive, insulin-releasing cells in a dish, offering hope for diabetes therapy.
Insights into Early Human Embryo Development
Researchers at Karolinska Institutet and the Ludwig Cancer Research in Stockholm have conducted a detailed molecular analysis of the embryo’s first week of development.
Boosting Gene Transfer Capabilities
A new and highly efficient method for gene transfer has been developed.
Liver-On-Chip Tracks Dynamics of Cellular Function
Hebrew University’s liver-on-chip platform is uniquely able to monitor metabolic changes indicating mitochondrial damage occurring at drug concentrations previously regarded as safe.
EU Project Aims to Cure Type 1 Diabetes
Researchers develop organoids from insulin-producing cells for transplantation.
Hope for Combating Muscular Dystrophy
Decoding a sugar molecule and identifying a mechanism linking it to MS could help in the development of therapy for the disease.
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