image: Fig. 1. Schematic diagram of the rapid FecBB detection using the Cas12a system.
Credit: Tingjie Wu, Xinjie Wang, Xiaolong Wang, et al.
BMPR1B is a gene that influences reproductive performance in sheep. Sheep populations that carry the FecBB mutation within this gene exhibit significantly higher lambing rates compared to wild-type populations. Therefore, screening for individuals carrying the FecBB mutation is crucial for effective sheep breeding programs.
A team of researchers from China establish a rapid, precise, and visualised on-site detection method for genotyping the prolific FecBB mutation in sheep. Their findings are published in the Journal of Integrative Agriculture.
"We combined the CRISPR/Cas12a system with the recombinase-polymerase amplification (RPA) technique and introduced an additional nucleotide mismatch on the amplification primers to form a Cas12a-recognised protospacer adjacent motif (PAM) sequence," explains senior and co-corresponding author Prof. Xinjie Wang from the Chinese Academy of Agricultural Sciences. "In addition, mismatches were introduced in CRISPR-derived RNA (crRNA) to enable naked-eye differentiation of the assay results."
Subsequently, the reserchers validated the accuracy of the method by examining additional blood samples from 56 sheep representing four breeds.
"The results of using our developed system were highly consistent with the Sanger sequencing," says co-corresponding author Prof. Xiaolong Wang from Northwest A&F University. "Overall, the CRISPR/Cas12a-based detection provides a rapid and more versatitle method for FecBB genotyping. It holds promise in enhancing efficiency in livestock breeding programmes for any single nucleotide mutations."
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Contact the author: Tingjie Wu, E-mail: tingjie.wu@nwafu.edu.cn; #Correspondence Xinjie Wang, E-mail: wangxinjie@caas.cn; Xiaolong Wang, Tel: +86-29-87091130, E-mail: xiaolongwang@nwafu.edu.cn
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Journal
Journal of Integrative Agriculture
Method of Research
Experimental study
Subject of Research
Animals
Article Title
Rapid on-site genotyping of the ovine prolific FecBB mutation using a CRISPR/Cas12a-based detection system
COI Statement
The authors declare that they have no conflict of interest.