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Rapid Genotyping of FecB Mutation in Sheep Using CRISPR-Cas12a Integrated with DNA Nanotree Biosensing Platform

文献类型： 外文期刊

作者： Pan, Dongxia 1 ; Mijit, Mukaddas 1 ; Wang, Hui 1 ; Sun, Chaoqun 1 ; Pingcuo, Bantan 2 ; Yu, Zhixue 1 ; Xiong, Benhai 1 ; Tang, Xiangfang 1 ;

作者机构： 1.CAAS, Inst Anim Sci, State Key Lab Anim Nutr & Feeding, Beijing 100193, Peoples R China

2.Tibet Autonomous Reg Acad Agr Sci, Inst Anim Husb & Vet, Lhasa 850004, Peoples R China

关键词： CRISPR-Cas12a; amplification retarded mutation system; FecB; Fe3O4@Au-Linker-Cy5-Nanotree; rapid detection

期刊名称：BIOMOLECULES （ 影响因子：4.8； 五年影响因子：5.6 ）

ISSN：

年卷期： 2025 年 15 卷 3 期

页码：

收录情况： SCI

摘要： The A-to-G mutation (FecB) in the BMPR1B gene is strongly linked to fertility in sheep, significantly increasing ovulation rates and litter sizes compared to wild-type populations. The rapid and reliable screening of the FecB gene is therefore critical for advancing sheep breeding programs. This study aimed to develop a fast and accurate method for detecting the FecB mutation and genotyping the gene to enhance sheep reproduction and productivity. To achieve this, we integrated the CRISPR-Cas12a system with an optimized amplification refractory mutation system (ARMS). A similar DNA origami technique-based fluorescence reporter nanotree structure was synthesized using gold nanomagnetic beads as carriers to amplify the fluorescence signal further. The resulting biosensing platform, termed CRISPR-ARMS, demonstrated excellent sensitivity for detecting FecB mutations, with a detection limit as low as 0.02 pmol. Therefore, this innovative approach shows great promise for single-base mutation detection and represents a pioneering tool for high-yield genetic screening.

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