|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.
|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.
|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.
|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.
|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.
|Assessment of the efficiency of encapsulation of a fluorescent drug using Nanoparticle Tracking Analysis|
Patrick Hole, Pierre Peotta, Roberto Santoliquido, Bob Carr
This poster outlines an example where the methodology of Nanoparticle Tracking Analysis (NTA) is used to characterise nanoparticles for drug delivery purposes.
|Bovine RNA-seq data analysis of liver and pituitary gland|
Pareek CS12, Smoczynski R12, Dziuba P12, Sikora M12, Golebiewski M2, Blaszczyk P12, Gelfand B3, Yaping F3, Kumar D3.
Two key applications of RNA-seq i.e., i) transcriptome read mapping to a reference genome and ii) SNP detections were investigated to analysis of bovine liver and pituitary gland transcriptome. Here, we have presented ONLY the obtained results of bovine pituitary gland.
|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.