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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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Quantification of Natural Killer Cell-Mediated Cytotixicity using Celigo Imaging Cytometry
Leo L. Chan, Srinivas S. Somanchi, Kelsey Rosbach, Dean A. Lee

Time-course tracking of % lysis eliminates multiple controls & the effect of non-uniform cell seeding in the final cytotoxicity calculation. The # of cells used is less than the cells needed for Release assays & Flow Cytometry. Flow cytometry & Release assays require a seeding density of 100K target cells increasing the number of effector cells to the millions. The visual observation of ADCC or CDC on the images can be convincing to conclude the functionality of antibodies or complements.

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A rapid 3D tumor spheroid analysis method using the Celigo Imaging Cytometry
Leo L. Chan, Scott Cribbes, Sarah Kessel, Olivier Dery, Catherine Swingler, Dmitry Kuksin, Tim Smith, Jean Qiu, Maria Vinci, Lisa Patterson, Sue Eccles

• Celigo Imaging Cytometer is a versatile and powerful tool for 3D tumorspheroid analysis
• Assays such as measuring optimal seeding density for forming tumorspheroids and viability measurement have been demonstrated
• Advanced assays such size measurement, growth inhibition, invasion into matrigel®, migration on to collagen and HUVEC, and tissue invasion that have been performed by manual microscope observation can now be easily quantified using the automated imaging cytometry method

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The Power Decoder simulator for the evaluation of pooled shRNA screen performance
Jesse Stombaugh, Abel Licon, Žaklina Strezoska, Joshua Stahl, Sarah Bael Anderson, Michael Banos, Anja van Brabant Smith, Amanda Birmingham, Annaleen Vermeulen

Power Decoder (written in R and Python) simulates shRNA pooled screening experiments in silico to allow for the estimation of a screen’s statistical power. Populations of shRNAs were engineered in such a way that the magnitude of depletion and enrichment was known, then using the negative binomial distribution, an in silico model was developed to successfully resemble data from an actual laboratory experiment.

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Knockdown of long noncoding RNAs in breast cancer
1 Jennii Luu, 2 Jesper Maag, 1 Yanny Handoko, 3 Richard Redvers, 3,4 Robin L. Anderson, 5 Maren M. Gross , 2 Marcel E. Dinger, and 1,3 Kaylene J. Simpson 1 Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre; 2 Genome Informatics, The Kinghorn Cancer Centre, The Garvan Institute of Medical Research; 3 Metastasis Research Laboratory, Peter MacCallum Cancer Centre, 4 Sir Peter MacCallum Department of Oncology, University of Melbourne;

RNAi global collaboration study using Lincode siRNA in a primary screen of tumor and nontumor breast cell lines. Hundreds of lncRNAs are found to affect viability and cell morphology of breast cancer. Presented at Keystone Symposia on Long Noncoding RNAs: From Evolution to Function, Mar 15 - Mar 20, 2015.

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Development of a novel xeno-free medium for feeder-free culture of human stem cells
Annand R; Okuda Y; Inamura M

A new xeno-free medium (ReproXFTM) has been developed for feeder-free culture of stem cells.

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Scientific News
Mitochondria Shown to Trigger Cell Ageing
An international team of scientists has for the first time shown that mitochondria, the batteries of the cells, are essential for ageing.
Cancer Cells Kill Off Healthy Neighbours
Cancer cells create space to grow by killing off surrounding healthy cells, according to UK researchers working with fruit flies.
Editing of Embryos Approved in the UK
The Human Fertilisation and Embryology Authority (HFEA) has approved a research application from the Francis Crick Institute to use new "gene editing" techniques on human embryos.
Microbes Take Their Vitamins
Scientists exploit organisms' needs in order to track 'vitamin mimics' in bacteria.
Machine Learning Uncovers Unknown Bacterial Features
Technique robustly identified characteristic gene expression patterns in response to antibiotics, low oxygen conditions.
CRISPR-Cas9 Gene Editing Advances Again
UC Berkeley researchers have made a major improvement in CRISPR-Cas9 technology that achieves an unprecedented success rate of 60 percent when replacing a short stretch of DNA with another.
Disrupting Cell’s Supply Chain Freezes Cancer Virus
When the cancer-causing Epstein-Barr virus moves into a B-cell of the human immune system, it tricks the cell into rapidly making more copies of itself, each of which will carry the virus.
Why Do Some Infections Persist?
In preparing for the possibility of an antibiotic onslaught, some bacterial cultures adopt an all-for-one/one-for-all strategy that would make a socialist proud, University of Vermont researchers have found.
ASCB: A CELLebration of Cell Biology
The last major congress of the year, ASCB is less a platform for launching new products, but one for confirming and consolidating the trends that have emerged over the past 12 months.
Squeezing Cells into Stem Cells
EPFL scientists have developed a new method that helps cells turn in usable stem cells. The new approach involves “squeezing” cells with a gel, and paves the way for large-scale production of stem cells for medical purposes.
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