Slide 1: Introduction
Introduce myself (Name, sophomore, Knockout Mice)
Slide 2: (what is a knockout mouse)
~A knockout mouse is a genetically engineered mouse.
~Researchers use the technique by “knocking out”/replacing the gene with an artificial piece of DNA. ~They do this so they could determine the specific function of the missing gene. ~Since humans are closely similar to mice, they can perform experiments on knockout mice and see the effects of the gene’s loss. ~This gives us a better understanding of a specific disease. ~This is a picture of a laboratory mouse where a gene is affecting hair growth had been knocked out (left), is shown next to a normal lab mouse.
Slide 3: (history)
~Mario R. Capecchi, Martin Evans and Oliver Smithies created the first known knockout mouse. ~They identified and separated the embryonic stem cells of the early embryo, the cell from which all cells of the adult organism are derived. ~He then established it in cell culture, modified it genetically and reintroduced it into foster mothers to generate a genetically modified offspring. ~In the picture (starting from the left, Oliver Smithies, Martin J. Evans and Mario R. Capecchi.
~Making a knockout mouse involves several steps. Takes about 2 yrs to get a mouse.
~In the first step of creating a knockout mouse, a “knockout” DNA construct is introduced into donor embryonic stem cells by electroporation. ~This construct is designed to introduce antibiotic resistance to cells if it gets in and recombines with the gene of interest.
~This is a picture of mouse embryonic stem cells growing on a bed of fibroblasts. These cells are put into a cuvette along with the DNA construct and loaded into the electroporator. ~This is an electroporator. It has a place for the cuvette to sit. The cells and DNA construct are then exposed to electricity and this changes the permeability of the cell membrane and DNA goes...