Children's Hospital of Eastern Ontario (CHEO) Research Institute

4 articles published in JoVE

• Genetics

  
  High-throughput DNA Extraction and Genotyping of 3dpf Zebrafish Larvae by Fin Clipping Ceres Kosuta^1,2, Kate Daniel¹, Devon L. Johnstone^1,2, Kevin Mongeon^1,2, Kevin Ban^1,2, Sophie LeBlanc¹, Stuart MacLeod¹, Karim Et-Tahiry², Marc Ekker², Alex MacKenzie¹, Izabella Pena^{1,2 1} Zebrafish have been used as reliable genetic model organisms in biomedical research, especially with the advent of gene-editing technologies. When larval phenotypes are expected, DNA extraction and genotype identification can be challenging. Here, we describe an efficient genotyping procedure for zebrafish larvae, by tail clipping, as early as 72-h post-fertilization.

• Cancer Research

  
  Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy Kristina J. Allan^1,2, Douglas J. Mahoney^1,4, Stephen D. Baird¹, Charles A. Lefebvre¹, David F. Stojdl^{1,2,3 1} Here we describe a protocol for employing high-throughput RNAi screening to uncover host targets that can be manipulated to enhance oncolytic virus therapy, specifically rhabodvirus and vaccinia virus therapy, but it can be readily adapted to other oncolytic virus platforms or for discovering host genes that modulate virus replication generally.

• Biology

  
  Isolation of CD146⁺ Resident Lung Mesenchymal Stromal Cells from Rat Lungs Jennifer J.P. Collins^1,2, Marius A. Möbius^1,3,4, Bernard Thébaud^{1,2,5 1}Sinclair Centre for Regenerative Medicine, Ottawa Hospital Research Institute, ²University of Ottawa, ³Department of Neonatology and Pediatric Critical Care Medicine, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, ⁴DFG Research Center and Cluster of Excellence for Regenerative Therapies (CRTD), Technische Universität, Dresden, ⁵ This protocol describes an isolation technique for obtaining primary lung resident mesenchymal stromal cells from rats, through the use of enzymatic digestion, density gradient separation, plastic adherence and CD146⁺ magnetic bead selection.

• Biology

  
  Identification of Kinase-substrate Pairs Using High Throughput Screening Courtney Reeks¹, Robert A. Screaton^{2,3 1} Protein phosphorylation is a central feature of how cells interpret and respond to information in their extracellular milieu. Here, we present a high throughput screening protocol using kinases purified from mammalian cells to rapidly identify kinases that phosphorylate a substrate(s) of interest.