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Selection, Genetic: Differential and non-random reproduction of different genotypes, operating to alter the gene frequencies within a population.

Detection of Residual Donor Erythroid Progenitor Cells after Hematopoietic Stem Cell Transplantation for Patients with Hemoglobinopathies

1Department of Pediatrics, Medical University Innsbruck, 2Department of Internal Medicine V (Hematology & Oncology), Medical University Innsbruck, 3Tyrolean Cancer Research Institute, 4Department of Pediatrics, Medical University Graz

JoVE 56002


 Developmental Biology

Phage Phenomics: Physiological Approaches to Characterize Novel Viral Proteins

1Department of Biology, San Diego State University, 2Computational Science Research Center, San Diego State University, 3Bioinformatics and Medical Informatics Research Center, San Diego State University, 4Department of Mathematics and Statistics, San Diego State University, 5Department of Computer Science, San Diego State University, 6Mathematics and Computer Science Division, Argonne National Laboratory, 7SPARC Committee, Broad Institute

JoVE 52854


 Immunology and Infection

Recombineering and Gene Targeting

JoVE 5553

One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based on stretches of similar or identical sequences. Together with gene targeting, which takes advantage of endogenous HR to alter an organism’s genome at a specific loci, HR-based cloning techniques have greatly improved the speed and efficacy of high-throughput genetic engineering.In this video, we introduce the principles of HR, as well as the basic components required to perform a recombineering experiment, including recombination-competent organisms and genomic libraries such as bacterial artificial chromosomes (BAC). We then walk through a protocol that uses recombineering to generate a gene-targeting vector that can ultimately be transfected into embryonic stem cells to generate a transgenic animal. Finally, several applications that highlight the utility and variety of recombineering techniques wi


 Genetics

Determining Immune System Suppression versus CNS Protection for Pharmacological Interventions in Autoimmune Demyelination

1Physical Medicine and Rehabilitation, University of Alabama at Birmingham, 2Department of Pathology, University of Alabama at Birmingham, 3Department of Neurobiology, University of Alabama at Birmingham, 4Center for Glial Biology and Medicine, University of Alabama at Birmingham

JoVE 54348


 Immunology and Infection

Measurement of Fronto-limbic Activity Using an Emotional Oddball Task in Children with Familial High Risk for Schizophrenia

1Department of Psychiatry, University of North Carolina at Chapel Hill School of Medicine, 2Duke-UNC Brain Imaging and Analysis Center, Duke University Medical Center, 3Curriculum in Neurobiology, University of North Carolina at Chapel Hill

JoVE 51484


 Behavior

Detection of Copy Number Alterations Using Single Cell Sequencing

1Koch Institute for Integrative Cancer Research, Department of Biology, Massachusetts Institute of Technology, 2Howard Hughes Medical Institute, 3Division of Health Sciences and Technology, Harvard Medical School, 4The Barbara K. Ostrom (1978) Bioinformatics and Computing Facility in the Swanson Biotechnology Center, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 5BioMicro Center, Department of Biology, Massachusetts Institute of Technology

JoVE 55143


 Genetics

Loss- and Gain-of-function Approach to Investigate Early Cell Fate Determinants in Preimplantation Mouse Embryos

1Department of Nanobiomedical Sciences and BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University, 2Insititute of Tissue Regeneration Engineering, Dankook University, 3Department of Biological Sciences, Ajou University, 4Department of Pharmacy, Sahmyook University, 5Department of Animal Biotechnology, Sahmyook University

JoVE 53696


 Developmental Biology

Preparation of Formalin-fixed Paraffin-embedded Tissue Cores for both RNA and DNA Extraction

1Department of Pathology & Molecular Medicine, Queen's University, 2Division of Cancer Biology & Genetics, Queen's Cancer Research Institute, Queen's University, 3Department of Surgery, Division of Urology, McGill University, 4Transformative Pathology Program, Ontario Institute for Cancer Research (OICR)

JoVE 54299


 Biology

Establishment of Cancer Stem Cell Cultures from Human Conventional Osteosarcoma

1Department of Surgery and Translational Medicine (DCMT), University of Florence, 2Neurofarba Department, University of Florence, 3Department of Traumatology and General Orthopedics, Azienda Ospedaliera Universitaria Careggi

JoVE 53884


 Cancer Research

Creating Objects and Object Categories for Studying Perception and Perceptual Learning

1Brain and Behavior Discovery Institute, Georgia Health Sciences University, 2Vision Discovery Institute, Georgia Health Sciences University, 3Department of Opthalmology, Georgia Health Sciences University, 4Intelligent Systems Laboratory, Palo Alto Research Center, 5Pattern Recognition Systems, Palo Alto Research Center, 6Department of Psychology, University of Minnesota

JoVE 3358


 Neuroscience

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