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Cattle with a precise, zygote-mediated deletion safely eliminate the major milk allergen beta-lactoglobulin

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posted on 2023-05-03, 18:01 authored by Jingwei WeiJingwei Wei, Stefan Wagner, Paul MacleanPaul Maclean, Brigid BrophyBrigid Brophy, Sally ColeSally Cole, Grant Smolenski, Daniel Carlson, Scott Fahrenkrug, David WellsDavid Wells, Goetz LaibleGoetz Laible
We applied zygote-mediated genome editing of the LGB gene to eliminate beta-lactoglobulin (BLG), a major allergen in cows’ milk. To efficiently generate LGB knockout cows, biopsied embryos were screened to transfer only appropriately modified embryos. Transfer of 13 pre-selected embryos into surrogate cows resulted in the birth three calves, one dying shortly after birth. Deep sequencing results confirmed conversion of the genotype from wild type to the edited nine bp deletion by more than 97% in the two male calves. The third calf, a healthy female, had in addition to the expected nine bp deletion (81 %), alleles with an in frame 21 bp deletion (< 17%) at the target site. While her milk was free of any mature BLG, we detected low levels of a BLG variant derived from the minor deletion allele. This demonstrated that the nine bp deletion genotype completely knocks out production of BLG. In addition, we showed that the LGB knockout animals are free of any TALEN-mediated off-target mutations or vector integration events using an unbiased whole genome analysis. Our study demonstrates the feasibility to generate precisely biallelically edited cattle, free of off-target mutations, using zygote-mediated editing for the safe production of hypoallergenic milk.


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Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made -


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Springer Nature

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Scientific Reports




Wei, J., Wagner, S., Maclean, P., Brophy, B., Cole, S., Smolenski, G., Carlson, D. F., Fahrenkrug, S. C., … Laible, G. (2018). Cattle with a precise, zygote-mediated deletion safely eliminate the major milk allergen beta-lactoglobulin. Scientific Reports, 8, 7661. doi:10.1038/s41598-018-25654-8

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