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| Cloning and Preliminary Functional Study of the OCA2 Gene in Oujiang Color Common Carp (Cyprinus carpio var. color) |
| ZENG Chun-Xiao1,2,3, KE Jing1,2,3, ZHANG Peng-Hui1,2,3, CHEN Hui-Fan1,2,3, CHEN Xiao-Wen1,2,3, WANG Jun1,2,3, WANG Cheng-Hui1,2,3,* |
1 Key Laboratory of Freshwater Aquatic Genetic Resources Certificated by the Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China;
2 National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;
3 Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China |
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Abstract The OCA2 (oculocutaneous albinism Ⅱ) gene is an important gene in melanin biosynthesis, and involved in pigmentation in the eye, skin, and hair colors of animals. Mutations in this gene could result in oculocutaneous albinism type Ⅱ in human. The black patchs on the body surface are key phenotypic characteristics in Oujiang color common carp (Cyprinus carpio var. color), however, the genetic basis and underlying mechanisms of their formation remain unclear. In this study, The high-generation selected strains of Oujiang color common carp was used to perform the cloning, expression and promoter transcriptional activity studies of the OCA2 gene, and gene knockout experiment was carried out by the CRISPR/Cas9 technology. The results revealed that the OCA2 gene consisted of 23 exons and 22 introns, encoding 834 amino acids with over 97% homology with the other fish species in Cyprinid family, which indicated highly structural and functional conservation. The qPCR analysis of the OCA2 gene showed that its expression levels were significantly higher in the eye and skin tissues in the whole white with black patch than those in the whole white color types in Oujiang color common carp. The dual-luciferase reporter assays indicated a complex regulatory mechanism in the promoter region of the OCA2 gene, and a key regulatory element during the 600~1 200 bp fragment before the start codon. CRISPR/Cas9-mediated knockout of exon and promoter regions in the OCA2 gene resulted in the numbers reduction by 70% and the areas decreasing by 40% of melanocytes from surface skin and eye tissues in the mutant. This study revealed that the OCA2 gene plays an important role in pigmentation in Oujiang color common carp.
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Received: 13 February 2025
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Corresponding Authors:
* Corresponding author, wangch@shou.edu.cn
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