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Thymidine Kinase: An enzyme that catalyzes the conversion of Atp and thymidine to Adp and thymidine 5'-phosphate. Deoxyuridine can also act as an acceptor and dGTP as a donor. (From Enzyme Nomenclature, 1992) EC 2.7.1.21.

In-vitro Mutagenesis

JoVE 10813

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

Genes can be randomly knocked out, or specific genes can be targeted. To knock out a particular gene, an engineered piece of DNA called a targeting vector is used to replace the normal gene, thereby inactivating it. Targeting vectors have sequences on each end that are identical—or homologous— to the sequences flanking each side of the gene of interest. These homologous sequences allow the targeting vector to replace the gene through homologous recombination—a process that occurs naturally between DNA with similar sequences during meiosis. The targeting vector is introduced into mouse embryonic stem cells in culture, using methods such as electroporation—use of electric pulses to temporarily create pores in the cell membrane. Typically, to identify cells where the vector has properly replaced the gene, it is designed to include a positive selection marker—such as the gene for neomycin resistance (NeoR)—between the homologous regions; and a negative selection marker—such as th

 Core: Biotechnology

Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy

1Children's Hospital of Eastern Ontario (CHEO) Research Institute, 2Department of Biology, Microbiology and Immunology, University of Ottawa, 3Department of Pediatrics, University of Ottawa, 4Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary

JoVE 56913

 Cancer Research

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

1Division of Gastroenterology and Liver Disease, University Hospitals, Digestive Health Research Institute, Case Western Reserve University, 2Case Cardiovascular Research Institute, School of Medicine, Case Western Reserve University, 3Department of Medicine, Harrington Discovery Institute, Harrington Heart and Vascular Institute, University Hospitals Case Medical Center, 4Department of Radiology, University Hospitals Case Medical Center, 5Department of Biology - Skeletal Research Center, Case Western Reserve University

JoVE 55367

 Medicine

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

1Center for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, 2Sino-British Research Centre for Molecular Oncology, National Center for International Research in Cell and Gene Therapy, Zhengzhou University, 3School of Basic Medical Sciences, Academy of Medical Sciences, Zhengzhou University

JoVE 54171

 Immunology and Infection

Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats

1II. Medizinische Klinik, Klinikum rechts der Isar der TU München, 2Department of Nuclear Medicine, Klinikum rechts der Isar der TU München, 3Division of Clinical Oncology, Hiroshima Prefectural Hospital, 4Center for Preclinical Research, Klinikum rechts der Isar der TU München, 5Institute for Diagnostic and Interventional Radiology, Klinikum rechts der Isar der TU München

JoVE 53757

 Medicine

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System

1Department of Neurosurgery, The University of Texas Health Science Center at Houston, 2Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 3The Senator Lloyd & B. A. Bentsen Center for Stroke Research, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 4Summer Research Program, Office of Educational Programs, The University of Texas Health Science Center at Houston, 5Department of Anesthesiology, Shengjing Hospital, China Medical University, 6Department of Oncology, Renji Hospital, Shanghai Jiaotong University School of Medicine, 7Biology Department, University of West Georgia

JoVE 52539

 Developmental Biology

Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling

1Department of Cell Biology, UT Southwestern Medical Center, 2National Institute of Neurological Disorders and Stroke, National Institute of Health, 3Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, 4Division of Genetics and Genomics, Boston Children's Hospital

JoVE 52307

 Medicine
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