A Novel Unstable Duplication Upstream of HAS2 Predisposes to a Breed-Defining Skin Phenotype and a Periodic Fever Syndrome in Chinese Shar-Pei Dogs
(A) Targeted resequencing of a 1.5 Mb region on chromosome 13 identified a duplication with on average 3.5–4.5X higher read coverage in two meatmouth Shar-Pei (black and red), compared to three control breeds (green, Standard Poodle; orange, Neapolitan Mastiff and purple, Pug). A shorter duplication was detected in the traditional Shar-Pei (blue). (B) The meatmouth duplication was determined to be 16.1 Kb long (CanFam 2.0 Chr13: 23,746,089–23,762,189) with both breakpoints located in repeats (a SINE and a LINE) and with an insertion of 7 bp separating different copies. The duplication in the traditional Shar-Pei overlapped the meatmouth duplication and was slightly shorter, 14.3 Kb long (CanFam 2.0 Chr13: 23,743,906–23,758,214). In this case the copies were separated by 1 bp but were still anchored in repeat motifs (c) Southern blot analysis using BsrGI digested gDNA from Shar-Pei and control breeds confirmed the existence of two duplication types in Shar-Pei. One meatmouth dog (lane 6) contained both duplication types. Individuals were classified as healthy (h) or as affected by Familial Shar-Pei Fever (f).