Versatile Xenopus tropicalis model with targeted integration of human BRAFV600E

• 成果类型: Article
• 署名作者: Ran, Rensen; Li, Lanxin; Chen, Peng; Li, Shuai; Wang, Peng; Zhu, Zhenpeng; Wang, Xiran; Chen, Yonglong; Hang, Jing; Liang, Weizheng
• 署名单位: Southern University of Science & Technology; Peking University; Xi'an Medical University; Guangdong Medical University; Hebei North University; Hebei North University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-11670
• DOI: 10.1073/pnas.2426981122
• 发表日期: 2025-09-30
• 关键词: pigment-epithelium; melanoma; cdkn2a; transdifferentiation; differentiation; regeneration; maintenance; laevis; SYSTEM; cells

摘要

Targeting exogenous gene integrations in animals often exhibits low efficiency, limiting the development of gene knock-in models. Theoretically, by screening founder generation individuals based on the cell phenotypes resulting from gene knock-ins and leveraging the high fecundity of animals, heritable descendants with targeted knock-ins can be efficiently generated. Therefore, we utilized the high fecundity of Xenopus tropicalis and easily observable pigment phenotypes to construct a BRAFV600E-targeted mitflocus knock-in model. Results indicated that this approach enabled efficient generation of BRAFV600E knock-in X. tropicalis and produced a versatile frog model. The BRAFV600E knock-in induced the transdifferentiation of RPE cells into retinal cells, resulting in a symmetric retinal structure in the eyes of these frogs. The transformation of RPE cells ultimately leads to these frogs becoming eyeless frogs, which serve as a tool for retinal regeneration research. Additionally, in eyeless frogs the BRAFV600E knock-in led to the abnormal proliferation of both melanocytes and xanthophores into melanocytic and xanthocytic nevi respectively. Consequently, eyeless frogs provide a model for studying abnormal pigment cell proliferation, offering a platform for investigating pigment cell nevus formation. Furthermore, the cdkn2b-knockout eyeless frogs serve as a valuable xanthophoroma model for tumor biology research. Overall, the BRAFV600E-targeted knock-in X. tropicalis not only represents a strategy for constructing gene knock-in animal models but also serves as a versatile tool for research in retinal regeneration and tumor biology.