8-Cell Injection

F0 generation mice fully derived from gene-targeted embryonic stem cells allowing immediate phenotypic analyses.

William T Poueymirou, Wojtek Auerbach, David Frendewey, Joseph F Hickey, Jennifer M Escaravage, Lakeisha Esau, Anthony T Dore, Sean Stevens, Niels C Adams, Melissa G Dominguez, Nicholas W Gale, George D Yancopoulos, Thomas M DeChiara& David M Valenzuela.

Nature Biotechnology, 2007. Jan. 25(1):91-99.

A useful approach for exploring gene function involves generating mutant mice from genetically modified embryonic stem (ES) cells. Recent advances in genetic engineering of ES cells have shifted the bottleneck in this process to the generation of mice. Conventional injections of ES cells into blastocyst hosts produce F0 generation chimeras that are only partially derived from ES cells, requiring additional breeding to obtain mutant mice that can be phenotyped. The tetraploid complementation approach directly yields mice that are almost entirely derived from ES cells, but it is inefficient, works only with certain hybrid ES cell lines and suffers from nonspecific lethality and abnormalities, complicating phenotypic analyses. Here we show that laser assisted injection of either inbred or hybrid ES cells into eight cell–stage embryos efficiently yields F0 generation mice that are fully ES cell–derived and healthy, exhibit 100% germline transmission and allow immediate phenotypic analysis, greatly accelerating gene function assignment.

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