Abstract
Most transgenic mice are generated by the direct microinjection of DNA fragments into the pronuclei of fertilized eggs. It has been generally assumed that the majority of integration events occur prior to the first round of chromosomal DNA replication (Palmiter and Brinster, 1986). In this study we have determined by comparison of PCR, Southern blot and transmission frequencies that at least 62% of integration events generate a mosaic (somatic and/or germline) G0 transgenic mouse. Furthermore, the statistical probability of transgene-containing cells segregating to the various early embryo lineages implies that this is probably an underestimate of the true mosaic frequency. Thus, the majority of DNA injected into fertilized mouse eggs integrates after the first round of chromosomal DNA replication, therefore most G0 transgenic mice are derived from a mosaic embryo.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brinster, R.L., Chen, H.Y., Traumbauer, M.E., Yagle, M.K. and Palmiter, R.D. (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjectin eggs.Proc. Natl Acad. Sci. USA 82, 4438–42.
Campbell, S.M., Rosen, J.M., Hennighausen, L.G., Strech-Jurk, U. and Sippel, A.E. (1984) Comparison of the whey acidic protein genes of the rat and mouse.Nucl. Acids Res. 12, 9685–7.
Clark, A.J., Archibald, A.L., McClenaghan, M., Simons, J.P., Whitelaw, C.B.A. and Wilmut, I. (1990) The germline manipulation of livestock: progress during the past five years.Proc. N.Z. Soc. Anim. Prod. 50, 167–79.
Covarrubias, L., Nishida, Y. and Mintz, B. (1986) Early postimplantation embryo lethality due to DNA rearrangements in transgenic mouse strain.Proc. Natl Acad. Sci. USA 83, 6020–4.
Gardner, R.L. (1978) The relationship between cell lineage and differentiation in the early mouse embryo. In Gehring, W.J. ed.,Genetic Mosaics and Cell Differentiation, pp. 211–41. Berlin: Springer-Verlag.
Hogan, B., Costantini, F. and Lacy, E. (1986)Manipulating the Mouse Embryo. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
Howlett, S.K. (1986) A set of proteins showing cell cycle dependent modification in the early mouse embryo.Cell 45, 387–96.
Markert, C.L. and Petters, R.M. (1978) Manufactured hexaparental mice show that adults are derived from three embryonic cells.Science 202, 56–8.
Mintz, B. (1970) Gene expression in allophenic mice. In Padykul, H. A. ed.,Control Mechanisms in the Expression of Cellular Phenotypes, pp. 15–42. New York: Academic Press.
Palmiter, R.D. and Brinster, R.L. (1986) Germ-line transformation of mice.Ann. Rev. Genet. 20, 465–99.
Pratt, H.P.M. (1987) Isolation, culture and manipulation of preimplantation mouse embryos. In Monk, M. ed.,Mammalian Development, pp. 13–42. Oxford: IRL Press.
Pursel, V.G., Miller, K.F., Bolt, D.J., Pinkert, C.A., Hammer, R.E., Palmiter, R.D. and Brinster, R.L. (1989) Insertion of growth hormone genes into pig embryos. In Heap, R.B., Prosser, C.G. and Lamming, G.E., eds.,Biotechnology in Growth Regulation, pp. 181–188. London: Butterworths.
Rexroad, C.E., Mayo, K., Bolt, D.J., Elsasser, T.H., Miller, K.F., Behringer, R.R., Palmiter, R.D. and Brinster, R.L. (1991) Transferrin- and albumin-directed expression of growthrelated peptides in transgenic sheep.J. Animal Sci. 69, 2995–3004.
Simons, J.P., Wilmut, I., Clark, A.J., Archibald, A.L., Bishop, J.O. and Lathe, R. (1988) Gene transfer into sheep.Bio Technology 6, 179–83.
Soriano, P. and Jaenisch, R. (1986) Retroviruses as probes for mammalian development: allocation of cells to the somatic and germ cell lineages.Cell 46, 19–29.
Telford, N.A., Watson, A.J. and Schultz, G.A. (1990) Transition from maternal to embryonic control in early mammalian development.Mol. Reprod. Dev. 26, 90–100.
Whitelaw, C.B.A., Archibald, A.L., Harris, S., McClenaghan, M., Simons, J.P., Springbett, A.J., Wallace, R. and Clark, A.J. (1990) Frequency of germline mosaicism in G0 transgenic mice.Mouse Genome 88, 114.
Whitelaw, C.B.A., Archibald, A.L., Harris, S., McClenaghan, M., Simons, J.P. and Clark, A.J. (1991) Targeting expression to the mammary gland: intronic sequences can enhance the efficiency of gene expression in transgenic mice.Transgenic Res. 1, 3–13.
Whitelaw, C.B.A., Harris, S., McClenaghan, M., Simons, J.P. and Clark, A.J. (1992) Position-independent expression of the ovine β-lactoglobulin gene in transgenic mice.Biochem. J. 286, 31–9.
Wilkie, T.M., Brinster, R.L. and Palmiter, R.D. (1986) Germline and somatic mosaicism in transgenic mice.Dev. Biol. 118, 9–18.
Wilmut, I., Hooper, M.L. and Simons, J.P. (1991) Genetic manipulation of mammals and its application in reproductive biology.J. Reprod. Fert. 92, 245–79.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Whitelaw, C.B.A., Springbett, A.J., Webster, J. et al. The majority of G0 transgenic mice are derived from mosaic embryos. Transgenic Research 2, 29–32 (1993). https://doi.org/10.1007/BF01977678
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01977678