caa a22 4500 397575513 CHVBK 20180308164855.0 cr unu---uuuuu 161202e199610 xx s 000 0 eng 10.1093/hmg/5.10.1523 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.10.1523 Full Genetic Rescue of Adenosine Deaminase-Deficient Mice Through Introduction of the Human Gene [Elektronische Daten] [Alexandra A. J. Migchielsen, Marco L. Breuer, Michael S. Hershfield, Dinko Valerio] We have shown recently that adenosine deaminase (ADA)-deficient mice die perinatally with severe liver cell degeneration. In addition to enzyme substitution, we report the restoration of viability through introduction of the human ADA gene. The ADA gene is subject to complex developmental and tissue-specific regulation. To include the cis-regulatory elements necessary for correct regulation of the human ADA gene, a large transgenic locus constituting the human ADA gene with 10 kb of 5′ and 4 kb of 3′ flanking sequences was generated by co-injection of two overlapping DNA fragments into murine zygotes. Probably as a result of extrachromosomal (homologous) recombination between the fragments, one of the two transgenic lines contained a reconstituted, functional human ADA gene. As in man, human ADA expression generally was low in these transgenic mice, but high in the thymus, spleen and gastro-duodenal part of the gut. Apparently, all cis-regulatory elements essential for a human expression pattern were incorporated in the transgene and were functional in the murine background. Similarly to man, the upper alimentary tract of the transgenic mice revealed low human ADA activity in contrast to extremely high levels of murine ADA. The human gene probably lacks the cis-regulatory elements that target high level murine ADA expression to the murine upper alimentary tract. ADA-deficient mice rescued by introduction of the human ADA transgene appeared histologically and immunologically normal. Apparently, human ADA can complement murine ADA in all tissues, even in the epithelium of the upper alimentary tract where human ADA activity is as much as 70-fold lower than murine ADA activity in wild-type mice. Clearly, the lethal phenotype of ADA-deficient mice is due to the absence of ADA. © 1996 Oxford University Press Migchielsen Alexandra A. J. Gene Therapy Section, Department of Medical Biochemistry, University of Leiden, Wassenaarseweg 72, P.O. Box 9503, 2300 RA Leiden, The Netherlands aut Breuer Marco L. Gene Therapy Section, Department of Medical Biochemistry, University of Leiden, Wassenaarseweg 72, P.O. Box 9503, 2300 RA Leiden, The Netherlands aut Hershfield Michael S. Department of Rheumatology and Immunology, Duke University Medical Center, Durham, NC 27710, USA aut Valerio Dinko IntroGene BV, Lange Kleiweg 151, 2280 GG Rijswijk, The Netherlands aut Human Molecular Genetics Oxford University Press 5/10(1996-10), 1523-1532 0964-6906 5:10<1523 1996 5 hmg https://doi.org/10.1093/hmg/5.10.1523 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.10.1523 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Migchielsen Alexandra A. J. Gene Therapy Section, Department of Medical Biochemistry, University of Leiden, Wassenaarseweg 72, P.O. Box 9503, 2300 RA Leiden, The Netherlands aut NATIONALLICENCE 700 1- Breuer Marco L. Gene Therapy Section, Department of Medical Biochemistry, University of Leiden, Wassenaarseweg 72, P.O. Box 9503, 2300 RA Leiden, The Netherlands aut NATIONALLICENCE 700 1- Hershfield Michael S. Department of Rheumatology and Immunology, Duke University Medical Center, Durham, NC 27710, USA aut NATIONALLICENCE 700 1- Valerio Dinko IntroGene BV, Lange Kleiweg 151, 2280 GG Rijswijk, The Netherlands aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/10(1996-10), 1523-1532 0964-6906 5:10<1523 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford