Open AccessManipulation of Mouse Embryonic Stem Cells for Knockout Mouse Production
Author(s) -
Limaye Advait,
Hall Bradford,
Kulkarni Ashok B.
Publication year - 2009
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/0471143030.cb1913s44
Subject(s) - embryonic stem cell , gene knockout , induced pluripotent stem cell , knockout mouse , biology , homologous recombination , gene targeting , microbiology and biotechnology , stem cell , conditional gene knockout , gene knockin , gene , genetics , cell culture , phenotype
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. Curr. Protoc. Cell Biol . 44:19.13.1‐19.13.24. © 2009 by John Wiley & Sons, Inc.