Frederick National Laboratory for Cancer Research 5 articles published in JoVE Genetics Optimization for Sequencing and Analysis of Degraded FFPE-RNA Samples Yelena Levin*1, Keyur Talsania*1,2, Bao Tran1, Jyoti Shetty1, Yongmei Zhao1,2, Monika Mehta1 1NCI CCR Sequencing Facility, Frederick National Laboratory for Cancer Research, 2Advanced Biomedical and Computational Sciences, Frederick National Laboratory for Cancer Research This method describes the steps to improve the quality and quantity of sequence data that can be obtained from formalin-fixed paraffin-embedded (FFPE) RNA samples. We describe the methodology to more accurately assess the quality of FFPE-RNA samples, prepare sequencing libraries, and analyze the data from FFPE-RNA samples. Biochemistry Fully Processed Recombinant KRAS4b: Isolating and Characterizing the Farnesylated and Methylated Protein Constance Agamasu1, Peter Frank1, Shelley Perkins1, Timothy Waybright1, Simon Messing1, William Gillette1, Andrew G. Stephen1 1NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research Prenylation is an important modification on peripheral membrane binding proteins. Insect cells can be manipulated to produce farnesylated and carboxymethylated KRAS4b in quantities that enable biophysical measurements of protein-protein and protein-lipid interactions Bioengineering Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate (LAL) Assays Barry W. Neun1, Marina A. Dobrovolskaia1 1Nanotechnology Characterization Lab., Cancer Research Technology Program, Frederick National Laboratory for Cancer Research sponsored by National Cancer Institute Detection of endotoxins in engineered nanomaterials represents one of the grand challenges in the field of nanomedicine. Here, we present a case study that describes the framework composed of three different LAL formats to estimate potential endotoxin contamination in nanoparticles. Chemistry Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications Matthew A. Ellis1, Giovanna Grandinetti2, Katye M. Fichter1 1Department of Chemistry, Missouri State University, 2Center for Molecular Microscopy, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research In this protocol, the synthesis of Cd-free InP/ZnS quantum dots (QDs) is detailed. InP-based QDs are gaining popularity due to the toxicity of Cd2+ ions that may be released through nanoparticle degradation. After synthesis, QDs are solubilized in water using an amphiphilic polymer for use in biomedical applications. Neuroscience A High Content Imaging Assay for Identification of Botulinum Neurotoxin Inhibitors Krishna P. Kota1,6, Veronica Soloveva2,6,7, Laura M. Wanner3,6, Glenn Gomba4,6, Erkan Kiris3,5,6, Rekha G. Panchal6, Christopher D. Kane2,6,7, Sina Bavari6 1Perkin Elmer Inc., 2Henry M. Jackson Foundation, 3The Geneva Foundation, 4ORISE, 5Frederick National Laboratory for Cancer Research, 6Division of Molecular and Translational Sciences, US Army Medical Research Institute of Infectious Diseases, 7DoD Biotechnology High Performance Computing Software Applications Institute (BHSAI), Telemedicine and Advanced Technology Research Center (TATRC), US Army Medical Research and Materiel Command (USAMRMC) Botulinum neurotoxin is one of the most potent toxins among Category-A biothreat agents, yet a post-exposure therapeutic is not available. The high content imaging approach is a powerful methodology for identifying novel inhibitors as it enables multiparameter screening using biologically relevant motor neurons, the primary target of this toxin.
