Patent ID: 7645917

Claim:
A method of producing a mouse homozygous for the mutated type II procollagen alpha-1 (Col2a-1) gene comprising the steps of: a) introducing into mouse embryonic stem cells a gene-targeting vector comprising a mutant Col2a-1 mouse gene which has exon 2 deleted but still comprises the remaining coding sequence for the wildtype Col2a-1 mouse gene; b) selecting for the mouse embryonic stem cells of step (a) wherein the endogenous Col2a-1 gene has been replaced by the mutant Col2a-1 mouse gene; c) introducing the mouse embryonic stem cells in step (b) into mouse blastocysts or aggregating the mouse embryonic stem cells selected in step (b) with a mouse morula; d) transplanting the mouse blastocysts or aggregated morulas of step (c) into a pseudopregnant mouse; e) allowing the blastocyst or morulas of step (d) to develop to term, thereby producing a chimeric founder mouse; f) crossing the chimeric founder mouse of step (e) with a wildtype mouse to produce a mouse heterozygous for the mutated Col2a-1 gene comprising in its genome a Col2a-1 gene that lacks exon 2 but still comprises the remaining coding sequence of the Col2a-1 gene and a wild-type Col2a-1 gene; and g) crossing mice heterozygous for the mutated Col2a-1 gene of step (f), thereby producing a mouse homozygous for the mutated Col2a-1 gene comprising in its genome a homozygous mutation of the Col2a-1 gene, wherein the mutated Col2a-1 gene lacks exon 2 but still comprises the remaining coding sequences of the Col2a-1 gene, wherein said mouse homozygous for the mutated Col2a-1 gene expresses a mutant Col2a-1 protein encoded by the mutant Col2a-1 gene, and displays multiple cardiovascular malformations similar to malformations present in human congenital hear defects, defects in morphogenesis of the heart similar to those associated with a loss of BMP, TGF-beta, and nodal signaling pathways including ventricular septal defect (VSD), patent ductus arteriosus (PDA), persistent truncus arteriosus (PTA), right ventricular hypertrophy, right aortic arch with or without anomalous origins of subclavian arteries, and a double outlet right ventricle (DORV) where aorta and pulmonary arteries arise wholly or in great part from the right ventricle, with associated VSD, transposition of great arteries (TOGA), anomalous coronary arteries, or endocardial cushion defect with primum atrial septal defect.