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Base pair editing of goat embryos: nonsense codon introgression into FGF5 to improve cashmere yield

By Guanwei Li, Shiwei Zhou, Chao Li, Bei Cai, Honghao Yu, Baohua Ma, Yu Huang, Yige Ding, Yao Liu, Qiang Ding, Chong He, Jiankui Zhou, Ying Wang, Guangxian Zhou, Yan Li, Yuan Yan, Jinlian Hua, Bjoern Petersen, Yu Jiang, Tad Sonstegard, Xingxu Huang, Yulin Chen, Xiaolong Wang

Posted 15 Jun 2018
bioRxiv DOI: 10.1101/348441

The ability to alter single bases without DNA double strand breaks provides a potential solution for multiplex editing of livestock genomes for quantitative traits. Here, we report using a single base editing system, Base Editor 3 (BE3), to induce nonsense codons (C-to-T transitions) at four target sites in caprine FGF5. All five progenies produced from microinjected single-cell embryos had alleles with a targeted nonsense mutation and yielded expected phenotypes. The effectiveness of BE3 to make single base changes varied considerably based on sgRNA design. Also, the rate of mosaicism differed between animals, target sites, and tissue type. PCR amplicon and whole genome resequencing analyses for off-target changes caused by BE3 were low at a genome-wide scale. This study provides first evidence of base editing in livestock, thus presenting a potentially better method to introgress complex human disease alleles into large animal models and provide genetic improvement of complex health and production traits in a single generation.

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