Non-meiotic allele introgression

A method of producing an infertile pig, the method comprising

• introducing into a pig cell a TALEN endonuclease or CRISPR composition comprising a CRISPR/Cas9 nuclease and a guide RNA, wherein the TALEN endonuclease or the guide RNA are designed to specifically bind a target sequence in exon 2 of an endogenous pig kiss-1 gene in the pig cell;
• introducing into the pig cell a homology-directed repair (HDR) template comprising a nucleic acid sequence encoding exon 2 of the endogenous kiss-1 gene or a portion thereof comprising a premature stop codon flanked by sequences homologous to exon 2 of the endogenous pig kiss-1 gene;
• wherein the TALEN endonuclease or the CRISPR/Cas9 nuclease, guided by the gRNA, cleave DNA in exon 2 of the endogenous pig kiss-1 gene in the pig cell and the HDR template inserts into the site of DNA cleavage by homologous recombination, whereby the premature stop codon is introduced into exon 2 of the endogenous pig kiss-1 gene and the endogenous pig kiss-1 gene is inactivated, thereby producing a gene-edited pig cell;
• transferring the gene-edited pig cell or DNA therefrom into an enucleated pig oocyte to obtain a gene-edited embryo;
• and
• implanting the gene-edited embryo into a surrogate female pig to produce a gene-edited pig whose endogenous kiss-1 gene is inactivated and the gene-edited pig is infertile.