Efficient in Vivo Manipulation of Mouse Genomic Sequences at the Zygote Stage

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1996 Jun 11

PMID 8650183

Citations 650

Authors

M Lakso

J G Pichel

J R Gorman

B Sauer

Y Okamoto

E Lee

F W Alt

H Westphal

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Evolutionary fingerprints of epithelial-to-mesenchymal transition.

Perelli L, Zhang L, Mangiameli S, Giannese F, Mahadevan K, Peng F Nature. 2025; .

PMID: 40044861 DOI: 10.1038/s41586-025-08671-2.

CTNNB1 syndrome mouse models.

Lainscek D, Forstneric V, Mirosevic S Mamm Genome. 2025; .

PMID: 39833474 DOI: 10.1007/s00335-025-10105-3.

Spontaneous mutation in 2310061I04Rik results in reduced expression of mitochondrial genes and impaired brain myelination.

Tsitsikov E, Phan K, Liu Y, Tsytsykova A, Paterno R, Sherry D PLoS One. 2024; 19(12):e0290487.

PMID: 39631040 PMC: 11617004. DOI: 10.1371/journal.pone.0290487.

A conditional smoothened (smo) allele on an inbred C57BL/6J genetic background has a hypomorphic smo mutant phenotype.

Houghtaling S, Gombart S, Ho T, Huang G, Beier D Dev Biol. 2024; 518():71-76.

PMID: 39603584 PMC: 11728190. DOI: 10.1016/j.ydbio.2024.11.010.

Directional ciliary beats across epithelia require Ccdc57-mediated coupling between axonemal orientation and basal body polarity.

Pan X, Fang C, Shen C, Li X, Xie L, Li L Nat Commun. 2024; 15(1):10249.

PMID: 39592607 PMC: 11599927. DOI: 10.1038/s41467-024-54766-1.

References

Mahon K, Chepelinsky A, Khillan J, Overbeek P, Piatigorsky J, Westphal H . Oncogenesis of the lens in transgenic mice. Science. 1987; 235(4796):1622-8. DOI: 10.1126/science.3029873. View

Schwenk F, Baron U, Rajewsky K . A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. Nucleic Acids Res. 1995; 23(24):5080-1. PMC: 307516. DOI: 10.1093/nar/23.24.5080. View

Thomas K, Capecchi M . Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987; 51(3):503-12. DOI: 10.1016/0092-8674(87)90646-5. View

Sauer B, Henderson N . Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc Natl Acad Sci U S A. 1988; 85(14):5166-70. PMC: 281709. DOI: 10.1073/pnas.85.14.5166. View

Mansour S, Thomas K, Capecchi M . Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature. 1988; 336(6197):348-52. DOI: 10.1038/336348a0. View

Dooley T, Miranda M, Jones N, DePamphilis M . Transactivation of the adenovirus EIIa promoter in the absence of adenovirus E1A protein is restricted to mouse oocytes and preimplantation embryos. Development. 1989; 107(4):945-56. DOI: 10.1242/dev.107.4.945. View

Sauer B, Henderson N . Targeted insertion of exogenous DNA into the eukaryotic genome by the Cre recombinase. New Biol. 1990; 2(5):441-9. View

Nakamura T, Mahon K, Miskin R, Dey A, Kuwabara T, Westphal H . Differentiation and oncogenesis: phenotypically distinct lens tumors in transgenic mice. New Biol. 1989; 1(2):193-204. View

Lakso M, Sauer B, Mosinger Jr B, Lee E, Manning R, Yu S . Targeted oncogene activation by site-specific recombination in transgenic mice. Proc Natl Acad Sci U S A. 1992; 89(14):6232-6. PMC: 49474. DOI: 10.1073/pnas.89.14.6232. View

10.

Orban P, Chui D, Marth J . Tissue- and site-specific DNA recombination in transgenic mice. Proc Natl Acad Sci U S A. 1992; 89(15):6861-5. PMC: 49604. DOI: 10.1073/pnas.89.15.6861. View

11.

Baubonis W, Sauer B . Genomic targeting with purified Cre recombinase. Nucleic Acids Res. 1993; 21(9):2025-9. PMC: 309460. DOI: 10.1093/nar/21.9.2025. View

12.

Gu H, Zou Y, Rajewsky K . Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting. Cell. 1993; 73(6):1155-64. DOI: 10.1016/0092-8674(93)90644-6. View

13.

Pichel J, Lakso M, Westphal H . Timing of SV40 oncogene activation by site-specific recombination determines subsequent tumor progression during murine lens development. Oncogene. 1993; 8(12):3333-42. View

14.

Gu H, Marth J, Orban P, Mossmann H, Rajewsky K . Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science. 1994; 265(5168):103-6. DOI: 10.1126/science.8016642. View

15.

Araki K, Araki M, Miyazaki J, Vassalli P . Site-specific recombination of a transgene in fertilized eggs by transient expression of Cre recombinase. Proc Natl Acad Sci U S A. 1995; 92(1):160-4. PMC: 42837. DOI: 10.1073/pnas.92.1.160. View

16.

Zou Y, Muller W, Gu H, Rajewsky K . Cre-loxP-mediated gene replacement: a mouse strain producing humanized antibodies. Curr Biol. 1994; 4(12):1099-103. DOI: 10.1016/s0960-9822(00)00248-7. View

17.

Kuhn R, Schwenk F, Aguet M, Rajewsky K . Inducible gene targeting in mice. Science. 1995; 269(5229):1427-9. DOI: 10.1126/science.7660125. View

18.

Kogan S, Doherty M, Gitschier J . An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. N Engl J Med. 1987; 317(16):985-90. DOI: 10.1056/NEJM198710153171603. View