"Genomic editing using CRISPR/Cas9 in cells and embryos to insert genes of interest in H11 locus of bovine genome."
Transgenic; Biotechnology; Animal Biology; Molecular Genetics; Genome Editing
Brazil has the largest commercial herd of cattle of the world with more than 210 million, in addition is currently the largest beef exporter. Through the development of this economic activity, the demand for the development of advanced technologies is also growing, including the production of genetically modified animals (GMA) that may address various problems from increased productivity to animal health and welfare. With this project, we intend to investigate the use of the CRISPR/Cas9 system as a model for establishing a genomic editing methodology that aims the direct insertion (knockin) of genes of biotechnological, agricultural or industrial interest in the H11 locus of the bovine (Bos taurus) genome. This study will analyze in bovine cells the capacity of the CRISPR/Cas9 system to insert expression cassettes of a reporter gene (eGFP) in a specific and safe site of the genome, called locus H11. In a second phase, the most effective conditions of the CRISPR/Cas9 system evaluated in the cells will be used for microinjection and/or electroporation of the CRISPR/Cas9 system directly into bovine zygotes. Our aim is directly editing the genome of bovine embryos, inserting genes of interest in a safe and predetermined location in the genome of this species of great zootechnical importance.