Polymerase chain reaction-based assays facilitate the breeding and study of mouse models of Klinefelter syndrome
Klinefelter syndrome (KS) is one of the most frequent genetic abnormalities and the leading genetic cause of non-obstructive azoospermia. The breeding of mouse models of KS and their study are essential to advance our knowledge of the pathologic mechanism. Karyotyping and fluorescence in situ hybridization are reliable methods for identifying chromosomal contents. However, technical issues associated with these methods can decrease the efficiency of breeding KS mouse models and limit studies that require rapid identification of target mice. To overcome these limitations, we developed three polymerase chain reaction-based assays to measure the specific genetic information, including the presence or absence of Sry, copy number of Amelx, and Xist RNA transcript levels. Through a combined analysis of the assay results, we can infer the karyotype of target mice. We confirmed the utility of our assays with the successful generation of KS mouse models. Our assays are rapid, inexpensive, high capacity, easy to perform, and require small amounts of sample. Therefore, they facilitate the breeding and study of KS mouse models and help advance our knowledge of the pathologic mechanism underlying KS.
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