|Experimental design considerations for efficient and specific gene knockin using a CRISPR-Cas9 for HDR with synthetic crRNA and tracrRNA|
Hidevaldo B. Machado, John A. Schiel, Maren Mayer-Gross, Eldon T. Chou, Melissa L. Kelley, Anja van Brabant Smith. Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Lafayette, CO 80026, USA
Precise genome engineering with CRISPR-Cas9 and single-stranded DNA oligo or double-stranded DNA plasmid donors via homology-directed repair (HDR).
|Exosomes promote survival in squamous head and neck cancer cells after ionizing radiation|
Lisa Mutschelknaus, Carsten Peters, Klaudia Winkler, Ramesh Yentrapalli, Theresa Heider, Michael J. Atkinson, Simone Moertl
Exosomes are important intercellular communicators in the acute radiation stress-response. These nanometer-sized extracellular vesicles confer protective signals to neighboring cells and therefore promote the tumorigenic and radioresistant phenotype of head and neck cancer cells.
|Super-resolution single molecule localization microscopy of the exocytotic machinery underlying insulin secretion.|
Deirdre M. Kavanagh, Alison Dun, Rory R. Duncan, and Colin Rickman.
Single molecule imaging of the tSNARE proteins involved in insulin secretion.
|A Synthetic CRISPR-Cas9 System for Homology-directed Repair|
John A. Schiel, Maren M. Gross, Emily M. Anderson*, Eldon T. Chou, Anja van Brabant Smith Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Lafayette, CO 80026, USA
Synthetic, dual-RNA-encoded Cas9 is used for precise homology-directed repair (HDR) gene engineering. Both short and long (GFP) inserts are covered.
|CRISPR-Cas9 genome editing utilizing chemically synthesized RNA|
Kaizhang He, Eldon Chou, Amanda Haas, Žaklina Strezoska, Melissa L. Kelley, and Anja van Brabant Smith Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Lafayette, CO 80026, USA
CRISPR-Cas9 gene editing using synthetic crRNA:tracrRNA or sgRNA is highly efficient and easy to use. Synthetic crRNA:tracrRNA is uniquely suited to in vitro and in vivo applications, in particular, DNA-free approach with Cas9 mRNA. Chemical synthesis of guide RNAs allows accurate and rapid production of arrayed crRNA libraries for high-confidence, loss-of-function screens.
|Deep Phenotyping - Harnessing Data Richness for Unsupervised High-Content Analysis|
Huang Dong, Wang Yi, Maciej Hermanowicz, Ke Yiping, Maja Choma, Lee Kee Khoon, Frederic Bard
Recognising the key challenges, we develop an end-to-end computational framework for HCA dubbed “Deep Phenotyping” that perform unsupervised analysis to leverage on the data richness for the discovery of unknown sub-phenotypes with minimal labeling cost.
|Assessment of the Anti-angiogenic Effect of VEGFR2 siRNA in Clonetics™ HUVEC using the Lonza 4D-Nucleofector™ System|
Srinivasan Kokatam1 , Kanchan Tiwari1 , Jenny Schroeder2 , Andrea Toell2 , Lubna Hussain3, Preeti Kapoor1
In the current study we have used siRNA targeting VEGFR2 as an example to study knockdown of VEGFR2 and subsequent inhibition of tube formation by HUVECs on Growth Factor Reduced Matrigel™ in a 96-well plate format. The same strategy can be used for screening and validating siRNA based inhibitors of the angiogenic process in vitro and thus could be of utility in anti-cancer screening strategies.
|Targeting Acute Pancreatitis by Small Molecule Inhibitors of Cyclophilin D|
M. Awais, E. Shore, R. Gibson, N. Kershaw, D. Latawiec, S. Pandalaneni, M.A. Javed, L. Wen, D.N. Criddle, N. Berry, L-Y. Lian, P. O’Neill, R. Sutton
Cyclophilin D (CypD) promotes opening of the mitochondrial permeability transition pore, a major contributor to acute pancreatitis. We are developing small molecule inhibitors of CypD as a possible treatment for AP and other conditions where the MPTP plays a role.
|The Pathogen Box: A Catalyst for Neglected Disease Drug Discovery|
Benoît Laleu*, Thomas Spangenberg, Wesley Van Voorhis, Angelique Doy, Dylan Pillai, Andreas Verras, Joie Garfunkle, Jeremy Burrows, Timothy Wells, Paul Willis
Modelled after the Malaria Box, the Pathogen Box contains ~400 diverse drug-like molecules active against neglected diseases of interest such as Chagas disease, cryptosporidiosis, hookworm, sleeping sickness, leishmaniasis, lymphatic filariasis, malaria, onchocerciasis, schistosomiasis, trichuriasis, tuberculosis.
|DNA-free CRISPR-Cas9 genome engineering in zebrafish|
Amanda Haas, Alex J. Blasky*, Rytis Prekeris*, John A. Schiel, Melissa L. Kelley, and Anja van Brabant Smith | Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite 100, Lafayette, CO 80026, USA *University of Colorado - Anschutz Medical Campus, Department of Cell & Developmental Biology, Denver, CO, USA
Poster describing the advantages of a DNA-free gene editing system and the application of this system in zebrafish.
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