Efficient in vivo manipulation of mouse genomic sequences at the zygote stage.

We describe a transgenic mouse line carrying the cre transgene under the control of the adenovirus EIIa promoter that targets expression of the Cre recombinase to the early mouse embryo. To assess the ability of this recombinase to excise loxP-flanked DNA sequences at early stages of development, we bred EIIa-cre transgenic mice to two different mouse lines carrying loxP-flanked target sequences: (i) a strain with a single gene-targeted neomycin resistance gene flanked by 1oxP sites and (ii) a transgenic line carrying multiple transgene copies with internal loxP sites. Mating either of these loxP-carrying mouse lines to EIIa-cre mice resulted in first generation progeny in which the loxP-flanked sequences had been efficiently deleted from all tissues tested, including the germ cells. Interbreeding of these first generation progeny resulted in efficient germ-line transmission of the deletion to subsequent generations. These results demonstrate a method by which loxP-flanked DNA sequences can be efficiently deleted in the early mouse embryo. Potential applications of this approach are discussed, including reduction of multicopy transgene loci to produce single-copy transgenic lines and introduction of a variety of subtle mutations into the line.

[1]  M Aguet,et al.  Inducible gene targeting in mice , 1995, Science.

[2]  J. Miyazaki,et al.  Site-specific recombination of a transgene in fertilized eggs by transient expression of Cre recombinase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[3]  K. Rajewsky,et al.  A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. , 1995, Nucleic acids research.

[4]  K. Rajewsky,et al.  Cre-loxP-mediated gene replacement: a mouse strain producing humanized antibodies , 1994, Current Biology.

[5]  K. Rajewsky,et al.  Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. , 1994, Science.

[6]  H. Westphal,et al.  Timing of SV40 oncogene activation by site-specific recombination determines subsequent tumor progression during murine lens development. , 1993, Oncogene.

[7]  K. Rajewsky,et al.  Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting , 1993, Cell.

[8]  B. Sauer,et al.  Genomic targeting with purified Cre recombinase. , 1993, Nucleic acids research.

[9]  J. Marth,et al.  Tissue- and site-specific DNA recombination in transgenic mice. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. Mosinger,et al.  Targeted oncogene activation by site-specific recombination in transgenic mice. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[11]  N. Henderson,et al.  Targeted insertion of exogenous DNA into the eukaryotic genome by the Cre recombinase. , 1990, The New biologist.

[12]  N. Jones,et al.  Transactivation of the adenovirus EIIa promoter in the absence of adenovirus E1A protein is restricted to mouse oocytes and preimplantation embryos. , 1989, Development.

[13]  T. Nakamura,et al.  Differentiation and oncogenesis: phenotypically distinct lens tumors in transgenic mice. , 1989, The New biologist.

[14]  Mario R. Capecchi,et al.  Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes , 1988, Nature.

[15]  N. Henderson,et al.  Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[16]  M. Capecchi,et al.  Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells , 1987, Cell.

[17]  J. Gitschier,et al.  An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. , 1987, The New England journal of medicine.

[18]  J. Piatigorsky,et al.  Oncogenesis of the lens in transgenic mice. , 1987, Science.

[19]  B. Hogan,et al.  Manipulating the mouse embryo: A laboratory manual , 1986 .