Dataset Information

CRISPR/Cas12a mediated knock-in of the Polled Celtic variant to produce a polled genotype in dairy cattle.

ABSTRACT: In modern livestock farming horned cattle pose an increased risk of injury for each other as well as for the farmers. Dehorning without anesthesia is associated with stress and pain for the calves and raises concerns regarding animal welfare. Naturally occurring structural variants causing polledness are known for most beef cattle but are rare within the dairy cattle population. The most common structural variant in beef cattle consists of a 202 base pair insertion-deletion (Polled Celtic variant). For the generation of polled offspring from a horned Holstein-Friesian bull, we isolated the Polled Celtic variant from the genome of an Angus cow and integrated it into the genome of fibroblasts taken from the horned bull using the CRISPR/Cas12a system (formerly Cpf1). Modified fibroblasts served as donor cells for somatic cell nuclear transfer and reconstructed embryos were transferred into synchronized recipients. One resulting pregnancy was terminated on day 90 of gestation for the examination of the fetus. Macroscopic and histological analyses proved a polled phenotype. The remaining pregnancy was carried to term and delivered one calf with a polled phenotype which died shortly after birth. In conclusion, we successfully demonstrated the practical application of CRISPR/Cas12a in farm animal breeding and husbandry.

SUBMITTER: Schuster F

PROVIDER: S-EPMC7419524 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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CRISPR/Cas12a mediated knock-in of the Polled Celtic variant to produce a polled genotype in dairy cattle.

Schuster Felix F Aldag Patrick P Frenzel Antje A Hadeler Klaus-Gerd KG Lucas-Hahn Andrea A Niemann Heiner H Petersen Björn B

Scientific reports 20200811 1

In modern livestock farming horned cattle pose an increased risk of injury for each other as well as for the farmers. Dehorning without anesthesia is associated with stress and pain for the calves and raises concerns regarding animal welfare. Naturally occurring structural variants causing polledness are known for most beef cattle but are rare within the dairy cattle population. The most common structural variant in beef cattle consists of a 202 base pair insertion-deletion (Polled Celtic varian ...[more]

PMID: 32782385

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Project description:Genetic modification of farm animals has not been well accepted by the public. Some modifications have the potential to improve animal welfare. One such example is the use of gene editing (i.e. CRISPR (clustered regularly interspaced short palindromic repeats)) to spread the naturally occurring POLLED gene, as these genetically hornless animals would not need to experience the painful procedures used to remove the horns or horn buds. The aim of the current study was to assess public attitudes regarding the use of GM to produce polled cattle. United States (US) citizens (n = 598), recruited via Amazon Mechanical Turk, were asked "Do you think genetically modifying cows to be hornless would be…", and responded using a 7-point Likert scale (1 = a very bad thing, 4 = neither good nor bad, 7 = a very good thing). Participants were then asked to indicate if they would be willing to consume products from these modified animals. We excluded 164 of the original 598 participants for not completing the survey, failing any of three attention check questions, or providing no or unintelligible qualitative responses. Respondents were then asked to provide a written statement explaining their answers; these reasons were subjected to qualitative analysis. Comparison of Likert scale ratings between two groups was done using the Wilcoxon rank-sum test, and comparisons between more than two groups were done using the Kruskal-Wallis rank test. More people responded that the modification would be good (Likert ≥ 5; 65.7%) than bad (Likert ≤ 3; 23.1%), and that they would be willing to consume products from these animals (Likert ≥ 5; 66.0%) versus not consume these products (Likert ≤ 3; 22.6%). Qualitative analysis of the text responses showed that participant reasoning was based on several themes including animal welfare, uncertainty about the technology, and worker well-being. In conclusion, many participants reported positive attitudes towards GM polled cattle; we suggest that people may be more likely to support GM technologies when these are perceived to benefit the animal.

Project description:BACKGROUND:Breeding genetically hornless, i.e. polled, cattle provides an animal welfare-friendly and non-invasive alternative to the dehorning of calves. However, the molecular regulation of the development of horns in cattle is still poorly understood. Studying genetic characters such as polledness and scurs, can provide valuable insights into this process. Scurs are hornlike formations that occur occasionally in a wide variety of sizes and forms as an unexpected phenotype when breeding polled cattle. METHODS:We present a unique dataset of 885 Holstein-Friesian cattle with polled parentage. The horn phenotype was carefully examined, and the phenotypic heterogeneity of the trait is described. Using a direct gene test for polledness, the polled genotype of the animals was determined. Subsequently, the existence of a putative scurs locus was investigated using high-density genotype data of a selected subset of 232 animals and two mapping approaches: mixed linear model-based association analyses and combined linkage disequilibrium and linkage analysis. RESULTS:The results of an exploratory data analysis indicated that the expression of scurs depends on age at phenotyping, sex and polled genotype. Scurs were more prevalent in males than in females. Moreover, homozygous polled animals did not express any pronounced scurs and we found that the Friesian polled allele suppresses the development of scurs more efficiently than the Celtic polled allele. Combined linkage and linkage disequilibrium mapping revealed four genome-wide significant loci that affect the development of scurs, one on BTA5 and three on BTA12. Moreover, suggestive associations were detected on BTA16, 18 and 23. The mixed linear model-based association analysis supports the results of the combined linkage and linkage disequilibrium analysis. None of the mapping approaches provided convincing evidence for a monogenic inheritance of scurs. CONCLUSIONS:Our results contradict the initial and still broadly accepted model for the inheritance of horns and scurs. We hypothesise an oligogenetic model to explain the development of scurs and polledness.

Dataset Information

CRISPR/Cas12a mediated knock-in of the Polled Celtic variant to produce a polled genotype in dairy cattle.

Publications

CRISPR/Cas12a mediated knock-in of the Polled Celtic variant to produce a polled genotype in dairy cattle.

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