Trans-splicing methods and compositions for generation of single sex offspring

A method of producing a single sex population of non-human vertebrate animals, the method comprising:

• crossing (i) a first non-human vertebrate animal having a first genotype comprising one or more sequence variants of a gene, and heterozygous allosomes, wherein one of the allosomes is modified to express one or more expression cassettes comprising a promoter, a nucleotide sequence, a splice site, an open reading frame encoding a transgenic protein, and a polyadenylation signal;
• with (ii) a second transgenic non-human vertebrate animal having a second genotype comprising a wildtype genome with homozygous allosomes;
• wherein a resulting progeny having a genotype comprising a wildtype gene and the allosome engineered to express the one or more transgenes is not viable;
• thereby creating a single sex population.

A method of producing a single sex population of non-human vertebrate animals, the method comprising:

• obtaining (i) a first non-human vertebrate animal comprising one or more sequence variants of an autosomal gene, and a modified allosome comprising one or more expression cassettes, wherein the one or more expression cassettes comprise the following elements in 5′ to 3′ orientation:
• a promoter;
• operatively linked thereto a nucleic acid sequence;
• a splice site;
• an open reading frame encoding a transgenic protein;
• and
• a polyadenylation signal;
• obtaining (ii) a second non-human vertebrate animal comprising a wildtype genome;
• and
• crossing the first non-human vertebrate animal and the second non-human vertebrate animals, wherein a resulting progeny comprising a wildtype gene and the modified allosome expressing the transgenic protein is not viable;
• thereby creating a single sex population.

A method of producing a single sex population of non-human vertebrate animals, the method comprising:

• crossing (i) a first non-human vertebrate animal having a first genotype comprising one or more nucleotide modifications in a sequence of an intron of a gene;
• and one or more expression cassettes comprising a promoter, a splice site, an open reading frame encoding a transgenic protein, and a polyadenylation signal, wherein the one or more nucleotide modifications in the sequence of the intron cannot splice to the splice site, and wherein the intron of the gene and the one or more expression cassettes are located on a single allosome;
• with (ii) a second transgenic non-human vertebrate animal having a second genotype comprising a wildtype sequence of the intron of the gene, wherein the wildtype sequence of the intron of the gene is capable of splicing to the splice site and homozygous allosomes;
• wherein a resulting progeny having a genotype comprising the wildtype sequence of the intron of the gene and the one or more expression cassettes is not viable.