|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.
|An HTS-Compatible Plate For Highly Miniaturized Cultures Of Primary Human Bronchial Epithelial Cells At Air-Liquid Interface|
Elizabeth Vu1, Eric Sorscher2, Robert Lowery1, Steven Hayes1
Primary human bronchial epithelial cells (HBE) cultured at air liquid interface (ALI) exhibit striking similarity to the in vivo situation, including both tissue architecture and ion channel functionality. Cultures of this type serve as a gold standard for predicting therapeutic activity in airway diseases such as cystic fibrosis.
|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.
|The Prestwick Chemical Library (R), A Valuable Tool for Screening|
Jean-Marie Contreras1, Christophe Morice1, Jean-Marc Simon1, Bruno Didier2, Marie-Louise Jung1 and Thierry Langer3
The Prestwick Chemical Library® (PCL) is Prestwick’s flagship product dedicated to screening. It is a collection of 1280 molecules comprising 100% approved drugs (FDA, EMEA and other agencies) selected for their high chemical and pharmacological diversity. The PCL was designed to reduce the risk of "low quality" hits and therefore the cost of the initial screening, and appears to be an efficient smart library for hit discovery. The PCL comes with an extended fully-annotated database.
|A proteomic analysis of p27kip1-binding proteins reveals a putative role in transcription regulation through RNA polymerase II interaction|
Juan Triviño Paredes1, Atilla Biçer1, Arnauld Besson2, Edurne Gallastegui1, Josep Maria Estanyol3, Maria Jesus Pujol1 and Oriol Bachs1
A proteomic analysis of p27kip1-binding proteins reveals a putative role in transcription regulation through RNA polymerase II interaction