|Improved Small RNA Library Preparation Workflow for Next-Generation Sequencing|
Sabrina Shore, Jordana Henderson, Anton McCaffrey, Gerald Zon, Richard Hogrefe
We describe an optimized small RNA NGS library prep workflow using chemically modified adapters which suppresses adapter dimers, allows for RNA inputs down to 1 ng and eliminates the need for a gel purification step, thus allowing full automation not previously possible.
|Specificity of highly potent miRNA inhibitors|
Barbara Robertson, Andrew Dalby, Yuriy Fedorov, Jon Karpilow, Anastasia Khvorova1, Devin Leake, Annaleen Vermeulen
miRNA inhibitors are invaluable tools for elucidating the roles of miRNAs. However, potent inhibitors may also affect other miRNAs. To understand the potential cross-reactivity of miRNA inhibitors, various miRNA inhibitor designs were systematically tested. We demonstrate that mismatches both within and outside the seed region of the miRNA interfere with inhibition. Our findings indicate that features important for natural miRNA target recognition are also important for inhibitor specificity.
|Targeting Cancer Stem Cell-Related miRNAs for Prostate Cancer Therapy|
ANSHIKA NIKITA SINGH, MEGHNA BARUAH, NEETI SHARMA
The poster focuses on the pivotal function of miRNAs in tumorigenesis by regulation of self renewal and apoptosis via cancer stem cell signalling pathways with special focus on their regulation of Epithelial to Mesenchymal transition in metastatic prostate cancer.
|siRNA Screening: Development of Hit Stratification Strategies|
Žaklina Strezoska, Annaleen Vermeulen, Emily M. Anderson, Anja Smith, Devin Leake
This poster compares different approaches to hit stratification and validation after an initial screen. Standard siRNA reagents deconvoluted from a pooled set of four were compared to a pooled set of four specificity enhanced reagents. High confidence hits were similar. To explore the validity of low confidence hits, a chimeric approach was used whereby a gene-specific seed sequence was introduced into a non-targeting siRNA scaffold. This work resulted in new hit stratification strategies.
|Identification of microRNA targets using microRNA modulation techniques and gene expression arrays|
Emily M. Anderson, Maren Mayer, Kevin Sullivan, Barbara Robertson, Žaklina Strezoska, Annaleen Vermeulen, and Devin Leake
By examining the overlap of messages down-regulated by miRNA mimics and up-regulated by miRNA inhibitors, we robustly identify miRNA-regulated messages, many of which have canonical seed matches and some which are not identied by standard target prediction programs.
|Specificity and functionality of microRNA inhibitors|
Barbara Robertson, Andrew Dalby, Jon Karpilow, Anastasia Khvorova, Devin Leake and Annaleen Vermeulen
Our findings indicate that features important for natural miRNA target recognition also appear to be important for inhibitor specificity. Understanding the specificity of inhibitors allows for better interpretation of inhibitor activity in endogenous systems.
|Alternative miRNA design for therapeutic RNAi applications|
Anja van Brabant Smith, Barb Robertson, Annaleen Vermeulen, Christina Yamada, Angela Reynolds, Anastasia Khvorova, Devin Leake
For in vivo applications, the design of miRNA inhibitors and miRNA mimics must be optimized for stability and potency. However, stabilized miRNA mimic molecules can lose functionality compared to standard miRNA mimic molecules due, in part, to the activity of the stabilized passenger strand acting as a miRNA inhibitor. We discuss how mismatches affect the activity of the stabilized miRNA mimics, perhaps by generating a passenger strand that is less functional as an inhibitor molecule.
|Integrating Fluorescent Carbon Nanodot Synthesis and Optical Detection of Methylmercury|
Carlos Bendicho, Isabel Costas-Mora, Vanesa Romero, Isela Lavilla
In the last years, a great interest toward development of optical nanoprobes has arisen, so fluorescent nanomaterials have been implemented in analytical systems for the detection of several species. In this work, a novel assay that integrates the synthesis of fluorescent carbon dots (CDs) and sensing within one step, for the fast, sensitive and selective detection of methylmercury is presented.
|Impact of Molecular Surface Charge on Electrical Impedance Spectroscopy Biosensing|
Y. Ram, T. Yoetz-Kopelman, A. Freeman and Y. Shacham-Diamand
Molecular surface charge was found to be the dominant parameter when monitoring protein binding events by Electrical Impedance Spectroscopy with a charged redox couple. A biosensing device was fabricated, and a physical model was derived to explain the results.
|A multiplexed amplicon sequencing technology for FFPE and circulating, cell-free DNA|
Laurie Kurihara, Catherine Couture, Julie Laliberte, Sukhinder Sandhu, Jonathan Irish, Tim Harkins and Vladimir Makarov
A novel amplicon approach allowing for hundreds of amplicons to be multiplexed in a single tube with a two workflow from sample to sequencer.
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