title: Generating double knockout mice to model genetic intervention for diabetic cardiomyopathy in humans.
abstract: Diabetes is a rapidly increasing disease that enhances the chances of heart failure twofold to fourfold (as compared to age and sex matched nondiabetics) and becomes a leading cause of morbidity and mortality. There are two broad classifications of diabetes: type1 diabetes (T1D) and type2 diabetes (T2D). Several mice models mimic both T1D and T2D in humans. However, the genetic intervention to ameliorate diabetic cardiomyopathy in these mice often requires creating double knockout (DKO). In order to assess the therapeutic potential of a gene, that specific gene is either overexpressed (transgenic expression) or abrogated (knockout) in the diabetic mice. If the genetic mice model for diabetes is used, it is necessary to create DKO with transgenic/knockout of the target gene to investigate the specific role of that gene in pathological cardiac remodeling in diabetics. One of the important genes involved in extracellular matrix (ECM) remodeling in diabetes is matrix metalloproteinase-9 (Mmp9). Mmp9 is a collagenase that remains latent in healthy hearts but induced in diabetic hearts. Activated Mmp9 degrades extracellular matrix (ECM) and increases matrix turnover causing cardiac fibrosis that leads to heart failure. Insulin2 mutant (Ins2+/-) Akita is a genetic model for T1D that becomes diabetic spontaneously at the age of 3-4 weeks and show robust hyperglycemia at the age of 10-12 weeks. It is a chronic model of T1D. In Ins2+/- Akita, Mmp9 is induced. To investigate the specific role of Mmp9 in diabetic hearts, it is necessary to create diabetic mice where Mmp9 gene is deleted. Here, we describe the method to generate Ins2+/-/Mmp9-/- (DKO) mice to determine whether the abrogation of Mmp9 ameliorates diabetic cardiomyopathy.
authors: Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, 985850 Nebraska Medical Center, Omaha, NE, 68198, USA.
authors: Vishalakshi
authors: V
authors: Chavali
authors: Shyam Sundar
authors: SS
authors: Nandi
authors: Shree Ram
authors: SR
authors: Singh
authors: Paras Kumar
authors: PK
authors: Mishra
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D058065:Diabetic Cardiomyopathies / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D004587:Electrophoresis, Agar Gel; D005260:Female; D055786:Gene Knockout Techniques / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D060005:Genotyping Techniques; D006801:Humans; D006824:Hybridization, Genetic; D006984:Hypertrophy; D007328:Insulin / Q000235:genetics; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D016133:Polymerase Chain Reaction; D014909:Wheat Germ Agglutinins / Q000378:metabolism
chemical_list: C561967:Ins2 protein, mouse; D007328:Insulin; D014909:Wheat Germ Agglutinins; D004247:DNA; D020780:Matrix Metalloproteinase 9