RNA guides genome engineering.

Laboratory of Cell and Gene Therapy, Center for Chronic Immunodeficiency, University Medical Center Freiburg, Freiburg, Germany.
Nature Biotechnology (Impact Factor: 39.08). 03/2013; 31(3):208-9. DOI: 10.1038/nbt.2527
Source: PubMed
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Genetically modified mice are powerful tools to investigate the molecular basis of many human diseases. Mice are, however, of limited value for preclinical studies, because they differ significantly from humans in size, general physiology, anatomy and lifespan. Considerable efforts are, thus, being made to develop alternative animal models for a range of human diseases. These promise powerful new resources that will aid the development of new diagnostics, medicines and medical procedures. Here, we provide a comprehensive review of genetically modified porcine models described in the scientific literature: various cancers, cystic fibrosis, Duchenne muscular dystrophy, autosomal polycystic kidney disease, Huntington's disease, spinal muscular atrophy, haemophilia A, X-linked severe combined immunodeficiency, retinitis pigmentosa, Stargardt disease, Alzheimer's disease, various forms of diabetes mellitus and cardiovascular diseases.
    Journal of applied genetics 11/2013; 55(1). DOI:10.1007/s13353-013-0182-9 · 1.90 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Only five years after the initiation of transgenic research in plants, gene targeting (GT) was achieved for the first time in tobacco. Unfortunately, the frequency of targeted integration via homologous recombination (HR) was so low in comparison to random integration that GT could not be established as a feasible technique in higher plants. It took another 25 years and great effort to develop the knowledge and tools necessary to overcome this challenge, at least for some plant species. In some cases, the overexpression of proteins involved in HR or the use of negative selectable markers improved GT to a certain extent. An effective solution to this problem was developed in 1996, when a sequence-specific endonuclease was used to induce a double-strand break (DSB) at the target locus. Thus, GT frequencies were enhanced dramatically. Thereafter, the main limitation was the absence of tools needed to induce DSBs at specific sites in the genome. Such tools became available with the development of zinc finger nucleases (ZFNs), and a breakthrough was achieved in 2005 when ZFNs were used to target a marker gene in tobacco. Subsequently, endogenous loci were targeted in maize, tobacco and Arabidopsis. Recently, our toolbox for genetic engineering has expanded with the addition of more types of site-specific endonucleases, meganucleases, transcription activator-like effector nucleases (TALENs) and the CRISPR/Cas system. We assume that targeted genome modifica-tions will become routine in the near future in crop plants using these nucleases along with the newly developed in planta GT technique.
    The International journal of developmental biology 01/2013; 57(6-7-8):629-637. DOI:10.1387/ijdb.130194hp · 2.57 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The molecular mechanisms underlying many human cancers are now reasonably well understood. The challenge now is to bridge the gap between laboratory and clinical oncology, so these accomplishments can be translated into practical benefits for human patients. While genetically modified mice have played a prominent role in basic research, they are less suitable for many preclinical studies. Other animals can provide important complementary resources to aid the development, validation and application of new medicines and procedures. Powerful methods of genetic engineering have now been extended to physiologically more relevant species, particularly the pig, opening the prospect of more representative, genetically defined, cancer models at human scale. We briefly review the field and outline our program to generate gene-targeted pigs carrying mutations in tumour suppressor genes and proto-oncogenes that replicate key lesions responsible for a variety of human cancers. We also highlight some important issues for the future development and usefulness of porcine cancer models.
    Transgenic Research 06/2013; 22(4). DOI:10.1007/s11248-013-9720-9 · 2.28 Impact Factor