Site-directed genome modification: derivatives of DNA-modifying enzymes as targeting tools

• Published in: Trends in biotechnology (Regular ed.) Vol. 23; no. 8; pp. 407 - 419
• Main Authors: Coates, Craig J., Kaminski, Joseph M., Summers, James B., Segal, David J., Miller, Andrew D., Kolb, Andreas F.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.08.2005; Elsevier Science; Elsevier Limited
• Subjects: Animals; Biological and medical sciences; Biotechnology; Deoxyribonucleic acid; DNA; DNA methylation; DNA Repair; Enzymes; Epigenetics; Exodeoxyribonucleases - genetics; Fundamental and applied biological sciences. Psychology; Gene Targeting - methods; Genetic engineering; Genetic Enhancement - methods; Genetic technics; Genetic Therapy - methods; Genomes; Genomics - methods; HIV; Human immunodeficiency virus; Humans; Leukemia; Methods. Procedures. Technologies; Miscellaneous; Mutagenesis; Mutagenesis, Site-Directed - genetics; Protein Engineering - methods; Proteins; Synthetic digonucleotides and genes. Sequencing; Site specificity; Targeting; Enzyme; Molecular integration; Review; Genome; Gene therapy; Application; Transgenesis
• ISSN: 0167-7799; 1879-3096
• DOI: 10.1016/j.tibtech.2005.06.009

The modification of mammalian genomes is an important goal in gene therapy and animal transgenesis. To generate stable genetic and biochemical changes, the therapeutic genes or transgenes need to be incorporated into the host genome. Ideally, the integration of the foreign gene should occur at sites that ensure their continual expression in the absence of any unwanted side effects on cellular metabolism. In this article, we discuss the opportunities provided by natural DNA-modifying enzymes, such as transposases, recombinases and integrases, to mediate the stable integration of foreign genes into host genomes. In addition, we discuss the approaches that have been taken to improve the efficiency and to modify the site-specificity of these enzymes.