Rapid Genotyping of <i>FecB</i> Mutation in Sheep Using CRISPR-Cas12a Integrated with DNA Nanotree Biosensing Platform
The A-to-G mutation (<i>FecB</i>) in the <i>BMPR1B</i> 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 <i>FecB</i> gene is therefore...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-02-01
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| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/15/3/315 |
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| Summary: | The A-to-G mutation (<i>FecB</i>) in the <i>BMPR1B</i> 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 <i>FecB</i> gene is therefore critical for advancing sheep breeding programs. This study aimed to develop a fast and accurate method for detecting the <i>FecB</i> 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 <i>FecB</i> 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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| ISSN: | 2218-273X |