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The first recorded knockout mouse was created by Mario R. Capecchi, Martin Evans, and Oliver Smithies in 1989, for which they were awarded the 2007 Nobel Prize in Physiology or Medicine. Aspects of the technology for generating knockout mice, and the mice themselves have been patented in many countries by private companies.
The International Knockout Mouse Consortium (IKMC) is a scientific endeavour to produce a collection of mouse embryonic stem cell lines that together lack every gene in the genome, and then to distribute the cells to scientific researchers to create knockout mice to study.
Conditional gene knockout is a technique used to eliminate a specific gene in a certain tissue, such as the liver. [1] [2] This technique is useful to study the role of individual genes in living organisms. It differs from traditional gene knockout because it targets specific genes at specific times rather than being deleted from beginning of life.
The Mouse Genetics Project (MGP) is a large-scale mutant mouse production and phenotyping programme aimed at identifying new model organisms of disease. [1] [2] [3] [4]Based at the Wellcome Trust Sanger Institute, the project uses knockout mice most of which were generated by the International Knockout Mouse Consortium.
The International Mouse Phenotyping Consortium (IMPC) is an international scientific endeavour to create and characterize the phenotype of 20,000 knockout mouse strains. [1] [2] [3] Launched in September 2011, [1] the consortium consists of over 15 research institutes across four continents with funding provided by the NIH, European national governments and the partner institutions.
Each stem cell contains one mutant gene copy and one 'wild-type' (normal) gene copy. The entire library is intended to mutate 13,000 genes in total. Of these 13000 mutant genes, 8000 mutations in mouse ES Cells are 'targeted': that is, the mutation which knocks out gene function is inserted precisely into the genome.
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Targeted gene knockout using CRISPR/Cas9 requires the use of a delivery system to introduce the sgRNA and Cas9 into the cell. Although a number of different delivery systems are potentially available for CRISPR, [ 37 ] [ 38 ] genome-wide loss-of-function screens are predominantly carried out using third generation lentiviral vectors.