Method for the rapid selection of homozygous primary cell lines for the production of transgenic animals by somatic cell nuclear transfer

A method for the accelerated production of transgenic animals homozygous for a selected trait comprising:

• transfecting a non-human mammalian cell-line with a given transgene construct containing at least one DNA encoding a desired gene;
• selecting a cell line(s) in which the desired gene has been inserted into the genome of that cell or cell-line;
• performing a first nuclear transfer procedure to generate a first transgenic animal heterzygous for the desired gene;
• characterizing the genetic composition of said first heterzygous transgenic animal;
• selecting cells homozygous for the desired transgene through the use of a selective agent;
• characterizing surviving cells using known molecular biology methods;
• and
• picking surviving cells or cell colonies cells for use in a second round of nuclear transfer or embryo transfer;
• and producing a second transgenic animal homozygous for a desired transgene.