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Gene Targeting: The integration of exogenous DNA into the genome of an organism at sites where its expression can be suitably controlled. This integration occurs as a result of homologous recombination.

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…

 Genetics

What is Genetic Engineering?

JoVE 10806

Genetic engineering is the process of modifying an organism’s DNA to introduce new, desirable traits. Many organisms, from bacteria to plants and animals, have been genetically modified for academic, medical, agricultural, and industrial purposes. While genetic engineering has definite benefits, ethical concerns surround modifying humans and our food supply.

Genetic engineering is possible because the genetic code—the way information is encoded by DNA—and the structure of DNA are universal among all life forms. As a result, an organism’s genetic code may be modified in several ways. The nucleotide sequence may be selectively edited by using techniques such as the CRISPR/Cas9 system. Known as the "molecular scissors," the CRISPR/Cas9 system is an innate, prokaryotic immune response that has been co-opted for editing genetic information. A gene may also be removed from an organism to create a “knockout,” or introduced to create a “knockin,” through a process called gene targeting. This method relies on homologous recombination—genetic exchange between DNA molecules that share an extended region with similar sequences—to modify an endogenous gene. Scientists can also insert a gene from one organism into the genome of another, resulting in a transgenic organism. Generally, DNA

 Core: Biotechnology

Genetic Engineering of Model Organisms

JoVE 5327

Transgenesis, or the use of genetic engineering to alter gene expression, is widely used in the field of developmental biology. Scientists use a number of approaches to alter the function of genes to understand their roles in developmental processes. This includes replacement of a gene with a nonfunctional copy, or adding a visualizable tag to a gene that allows the resultant fusion protein to …

 Developmental Biology

Genome Editing

JoVE 5554

A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be laborious and only works in certain organisms. Recent advances have led to the development of “genome editing”, which works by inducing double-strand breaks in DNA using engineered nuclease enzymes guided to target…

 Genetics

An Overview of Genetic Engineering

JoVE 5552

Genetic engineering – the process of purposefully altering an organism’s DNA – has been used to create powerful research tools and model organisms, and has also seen many agricultural applications. However, in order to engineer traits to tackle complex agricultural problems such as stress tolerance, or to realize the promise of gene therapy for treating…

 Genetics

Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing

1School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, 2Department of Animal Wealth Development, Faculty of Veterinary Medicine, Suez Canal University, 3Anatomy and Embryology Department, Faculty of Veterinary Medicine, Cairo University, 4Department of Molecular Physiology and Biophysics, Vanderbilt University, 5Life Science Institute, University of Michigan

JoVE 56275

 Genetics

CRISPR-mediated Loss of Function Analysis in Cerebellar Granule Cells Using In Utero Electroporation-based Gene Transfer

1Division of Molecular Neurogenetics, German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, 2Division of Molecular Genetics, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 3Hopp-Children's Cancer Center at the NCT Heidelberg (KiTZ), 4Division of Pediatric Neurooncology, German Cancer Research Center (DKFZ), and German Cancer Consortium (DKTK), 5Department of Pediatric Hematology and Oncology, Heidelberg University Hospital

JoVE 57311

 Developmental Biology

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

1Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 2College of Life Sciences, University of Chinese Academy of Sciences, 3School of Life Science and Technology, Shanghai Tech University, 4Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences

JoVE 56844

 Genetics

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii

1Laboratory of Molecular Biology, Department of Plant Sciences, Wageningen University & Research, 2Center of Technology for Agricultural Production, Agency for the Assessment and Application of Technology (BPPT), 3Department of Ecological Science, Faculty of Earth and Life Sciences, Vrije Universiteit Amsterdam, 4Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing University of Agriculture

JoVE 59971

 Genetics

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development

1Department of Biochemistry, University of Illinois at Urbana-Champaign, 2Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 3Neuroscience Program, University of Illinois at Urbana-Champaign, 4Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign

JoVE 55823

 Developmental Biology

Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

1Lab of Animal Models and Functional Genomics (LAMFG), The Key Laboratory of Animal Vaccine & Protein Engineering, College of Veterinary Medicine, Hunan Agricultural University (HUNAU), 2Department of Pathology, Georgetown University Medical School, 3College of Food Science and Technology, Hunan Agricultural University (HUNAU), 4Lab of Molecular Cardiology (LMC), National Heart, Lung, and Blood Institute (NHLBI)/National Institutes of Health (NIH), 5Transgenic Core, National Heart, Lung, and Blood Institute (NHLBI)/National Institutes of Health (NIH)

JoVE 58467

 Genetics

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9

1Stem Cells & Regenerative Medicine Center, Baylor College of Medicine, 2Center for Cell and Gene Therapy, Baylor College of Medicine, 3Centro di Ricerca Emato-Oncologica (CREO), University of Perugia, 4Department of Molecular & Human Genetics, Baylor College of Medicine, 5Texas Children's Hospital & Houston Methodist Hospital

JoVE 57278

 Genetics

Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex

1Department of Animal, Dairy, and Veterinary Sciences, Utah State University, 2National Centre for International Research in Cell and Gene Therapy, Sino-British Research Centre, School of Basic Medical Sciences, Zhengzhou University, 3Department of Animal Science Division of Applied Life Science (BK21 Plus), Gyeongsang National University, 4Institute of Agriculture and Life Science, Gyeongsang National University, 5Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London

JoVE 56263

 Bioengineering
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