Abstract
Worldwide, congenital deafness and pigmentation disorders impact millions with their diverse manifestations, and among these genetic conditions, mutations in the Microphthalmia-associated transcription factor (MITF: OMIM#156845) gene are notable for their profound effects on melanocyte development and auditory functions. This study reports a novel porcupine model exhibiting spontaneous deafness and pigmentation abnormalities reminiscent of human Waardenburg Syndrome Type 2 (WS2: OMIM#193510). Through phenotypic characterization, including coat color, skin, eye morphology, and auditory brainstem response (ABR) assessments, we identified hypopigmentation and complete deafness in mutant porcupines. To pinpoint the genetic basis, a breeding program was established, and Bulk Segregant Analysis (BSA) combined with RNA sequencing was conducted. Primers based on the identified candidate genes were designed for PCR amplification, followed by verification through Sanger sequencing. Through BSA analysis, we identified a total of 88 SNP and 336 InDel candidate sites. By annotating the Mitf gene, we obtained four unique transcript sequences. The SNP and InDel sites within the porcupine Mitf gene sequence, identified through BSA screening, were analyzed in conjunction with the gene's annotation results. This analysis revealed a specific mutation site, Mitf c.875_877delGAA p. (Arg217del), which was subsequently verified by Sanger sequencing. This naturally occurring Mitf mutation in porcupines provides a valuable model for studying the mechanisms underlying WS2 and exploring potential therapeutic strategies for deafness and pigmentation-related disorders.