Manipulation of mouse embryonic stem cells for knockout mouse production.
The establishment of mouse embryonic stem (ES) cell lines has allowed for the gene?ration of the knockout mouse. ES cells that are genetically altered in culture can then be manipulated to derive a whole mouse containing the desired mutation. To successfully generate a knockout mouse, however, the ES cells must be carefully cultivated in a pluripotent state throughout the gene-targeting experiment. This unit describes detailed step-by-step protocols, reagents, equipment, and strategies needed for the successful generation of gene knockout embryonic stem cells using homologous recombination technologies.
Limaye
Advait
A
Department of Health and Human Services, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
Hall
Bradford
B
Kulkarni
Ashok B
AB
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D055786:Gene Knockout Techniques / Q000379:methods*; D051379:Mice / Q000235:genetics*; D018345:Mice, Knockout; D011995:Recombination, Genetic