Open AccessGeneration of Gene Knockout Mice by ES Cell Microinjection
Author(s) -
Longenecker Glenn,
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.cb1914s44
Subject(s) - microinjection , blastocyst , gene knockout , knockout mouse , embryo , germline , biology , andrology , microbiology and biotechnology , gene , conditional gene knockout , embryonic stem cell , immunology , genetics , embryogenesis , medicine , phenotype
This unit describes protocols used in the production of chimeric mice that are then used for the generation of gene knockout mice. These protocols include the collection of blastocyst embryos, ES cell injection, and uterine transfer of injected blastocysts. Support protocols for the superovulation of blastocyst donor mice, generation of pseudopregnant recipients, fabrication of glass pipets for embryo and cell manipulations, and generation of germline mice are also included. Practical tips and solutions are mentioned to help troubleshoot problems that may occur. Curr. Protoc. Cell Biol . 44:19.14.1.‐19.14.36. © 2009 by John Wiley & Sons, Inc.