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Imprint
Sex determination in pigs by CRISPR/Cas (Gene-Editing Pigs)
Project
Project code: FLI-ING-08-HM-0003
Contract period: 01.04.2017
- 30.10.2020
Purpose of research: Experimental development
The aim of the project is to determine the possibility to influence the sex of pig offspring by CRISPR/Cas and to determine the function of the SRY gene in pigs. In a first step, we characterize the function of the Y-chromosomal porcine SRY gene in the process of sex determination. Therefore, a SRY-specific CRISPR/Cas expression vector will be transfected into male porcine fibroblasts and the mutation rate will be detected by using a T7En1 assay. Genetically modified fibroblasts will subsequently be used as donor cells in somatic cell nuclear transfer to produce cloned offspring and to determine the ratio of the sexes. Usually, the offspring should have a male phenotype as they originated from a male cell line. If SRY has the same sex determining in the as described in mice and rabbits, we should obtain male offspring with a female phenotype of the external and internal genital organs. In a second step, we will integrate a CRISPR/Cas expression vector onto the Y-chromosome targeting several essential sites of the Y-chromosome. This vector will carry a spermatogenesis-specific promoter driving the expression of Cas9. This approach is aiming at generating a complete destruction of the Y-chromosome during spermatogenesis to let only X-chromosomal sperm to develop. Mating of such a genetically modified boar would lead to only female offspring. This project is mainly addressing basic research but also tackles a huge problem occurring with the prohibition of the anesthesia-free castration of piglets under the age of 8 days in the EU from 2019.
Section overview
Subjects
- Animal breeding
