title: Potential molecular consequences of transgene integration: The R6/2 mouse example.
abstract: Integration of exogenous DNA into a host genome represents an important route to generate animal and cellular models for exploration into human disease and therapeutic development. In most models, little is known concerning structural integrity of the transgene, precise site of integration, or its impact on the host genome. We previously used whole-genome and targeted sequencing approaches to reconstruct transgene structure and integration sites in models of Huntington's disease, revealing complex structural rearrangements that can result from transgenesis. Here, we demonstrate in the R6/2 mouse, a widely used Huntington's disease model, that integration of a rearranged transgene with coincident deletion of 5,444 bp of host genome within the gene Gm12695 has striking molecular consequences. Gm12695, the function of which is unknown, is normally expressed at negligible levels in mouse brain, but transgene integration has resulted in cortical expression of a partial fragment (exons 8-11) 3' to the transgene integration site in R6/2. This transcript shows significant expression among the extensive network of differentially expressed genes associated with this model, including synaptic transmission, cell signalling and transcription. These data illustrate the value of sequence-level resolution of transgene insertions and transcription analysis to inform phenotypic characterization of transgenic models utilized in therapeutic research.
authors.affiliation: Centre for Brain Research, School of Biological Sciences, The University of Auckland 1010, New Zealand.
authors.forename: Jessie C
authors.initials: JC
authors.lastname: Jacobsen
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.|Program in Medical and Population Genetics, Broad Institute of M.I.T and Harvard, Cambridge, Massachusetts 02143, USA.
authors.forename: Serkan
authors.initials: S
authors.lastname: Erdin
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.|McDonnell Genome Institute, Washington University School of Medicine, St. Louis, Missouri 63108, USA.
authors.forename: Colby
authors.initials: C
authors.lastname: Chiang
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
authors.forename: Carrie
authors.initials: C
authors.lastname: Hanscom
authors.affiliation: Centre for Brain Research, School of Biological Sciences, The University of Auckland 1010, New Zealand.
authors.forename: Renee R
authors.initials: RR
authors.lastname: Handley
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Barker
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
authors.forename: Alex
authors.initials: A
authors.lastname: Stortchevoi
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
authors.forename: Ian
authors.initials: I
authors.lastname: Blumenthal
authors.affiliation: Centre for Brain Research, School of Biological Sciences, The University of Auckland 1010, New Zealand.
authors.forename: Suzanne J
authors.initials: SJ
authors.lastname: Reid
authors.affiliation: Centre for Brain Research, School of Biological Sciences, The University of Auckland 1010, New Zealand.
authors.forename: Russell G
authors.initials: RG
authors.lastname: Snell
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.|Program in Medical and Population Genetics, Broad Institute of M.I.T and Harvard, Cambridge, Massachusetts 02143, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts 02115 USA.
authors.forename: Marcy E
authors.initials: ME
authors.lastname: MacDonald
authors.affiliation: Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3DY, United Kingdom.
authors.forename: A Jennifer
authors.initials: AJ
authors.lastname: Morton
authors.affiliation: Department of Psychiatry, McGill University, Montreal, Quebec ON H4H 1R3, Canada.
authors.forename: Carl
authors.initials: C
authors.lastname: Ernst
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.|Program in Medical and Population Genetics, Broad Institute of M.I.T and Harvard, Cambridge, Massachusetts 02143, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115 USA.
authors.forename: James F
authors.initials: JF
authors.lastname: Gusella
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.|Program in Medical and Population Genetics, Broad Institute of M.I.T and Harvard, Cambridge, Massachusetts 02143, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts 02115 USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, 02114 USA.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Talkowski
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D015321:Gene Rearrangement; D006801:Humans; D006816:Huntington Disease / Q000473:pathology*; D051379:Mice; D008822:Mice, Transgenic; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion