Abstract
Over the past decades, transgenic mice have become a valuable tool in investigating gene expression, regulation, and function. Mouse models have been established for various human diseases to determine gene expression patterns and to elucidate gene function in tissues and organ systems. However, the outcome of a particular transgenic experiment is affected by a variety of factors that are only partly understood. Expression of transgenes may occur at ectopic sites, mouse lines generated might not express the transgene at all, or expression levels may decrease with increasing number of generations (see Note 1). Some of these difficulties can be solved by carefully planning the experiment. Thus the following chapter will not only deal with practical aspects of the microinjection but intends to provide guidelines for setting up a transgenic experiment.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ryding, A. D., Sharp, M. G., and Mullins, J. J. (2001) Conditional transgenic technologies. J. Endocrinol. 171, 1ā14.
McBurney, M. W., Staines, W. A., Boekelheide, K., Parry, D., Jardine, K., and Pickavance, L. (1994) Murine PGK-1 promoter drives widespread but not uniform expression in transgenic mice. Dev. Dyn. 200, 278ā293.
Ikawa, M., Yamada, S., Nakanishi, T., and Okabe, M. (1998) āGreen miceā and their potential usage in biological research. FEBS Lett. 430, 83ā87.
Kisseberth, W. C, Brettingen, N. T., Lohse, J. K., and Sandgren, E. P. (1999) Ubiquitous expression of marker transgenes in mice and rats. Dev. Biol. 214, 128ā138.
Potts, W., Tucker, D., Wood, H., and Martin, C. (2000) Chicken beta-globin 5ā²HS4 insulators function to reduce variability in transgenic founder mice. Biochem. Biophys. Res. Commun. 273, 1015ā1018.
Cranston, A., Dong, C, Howcroft, J., and Clark, A. J. (2001) Chromosomal sequences flanking an efficiently expressed transgene dramatically enhance its expression. Gene 269, 217ā225.
Jang, S. K., Krausslich, H. G., Nicklin, M. J., Duke, G. M., Palmenberg, A. C, and Wimmer, E. (1988) A segment of the 5ā² nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation. J. Virol. 62, 2636ā2643.
Takeuchi, T., Yamazaki, Y., Katoh-Fukui, Y, et al. (1995) Gene trap capture of a novel mouse gene, jumonji, required for neural tube formation. Genes Dev. 9, 1211ā1222.
Li, X., Wang, W., and Lufkin, T. (1997) Dicistronic LacZ and alkaline phosphatase reporter constructs permit simultaneous histological analysis of expression from multiple transgenes. Biotechniques 23, 874ā878, 880, 882.
Previtali, S. C, Quattrini, A., Fasolini, M., et al. (2000) Epitope-tagged P(0) glycoprotein causes Charcot-Marie-Tooth-like neuropathy in transgenic mice. J. Cell Biol. 151, 1035ā1046.
Oster-Granite, M. L., McPhie, D. L., Greenan, J., and Neve, R. L. (1996) Age-dependent neuronal and synaptic degeneration in mice transgenic for the C terminus of the amyloid precursor protein. J. Neurosci. 16, 6732ā6741.
Wight, P. A., Duchala, C. S., Readhead, C, and Macklin, W. B. (1993) A myelin proteolipid protein-LacZ fusion protein is developmentally regulated and targeted to the myelin membrane in transgenic mice. J. Cell Biol. 123, 443ā454.
Young, W. S. III, Iacangelo, A., Luo, X. Z., King, C, Duncan, K., and Ginns, E. I. (1999) Transgenic expression of green fluorescent protein in mouse oxytocin neurones. J. Neuroendocrinol. 11, 935ā939.
Sambrook, J., and Russell, D. W., eds. (2001) Molecular cloning: A laboratory manual, 3ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Hogan, B., Beddington, R., Costantini, F., and Lacy, E., eds. (1994) Manipulating the mouse embryo: A laboratory manual, 2ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Boado, R. J., Li, J. Y, Nagaya, M., Zhang, C, and Pardridge, W. M. (1999) Selective expression of the large neutral amino acid transporter at the blood-brain barrier. Proc. Natl. Acad. Sci. USA 96, 12,079ā12,084.
Qin, Y., and Sato, T. N. (1995) Mouse multidrug resistance 1a/3 gene is the earliest known endothelial cell differentiation marker during blood-brain barrier development. Dev. Dyn. 202, 172ā180.
Li, J. Y., Boado, R. J., and Pardridge, W. M. (2001) Blood-brain barrier genomics. J. Cereb. Blood Flow Metab. 21, 61ā68.
Lamb, B. T., and Gearhart, J. D. (1995) YAC transgenics and the study of genetics and human disease. Curr. Opin. Genet. Dev. 5, 342ā348.
Peterson, K. R., Clegg, C. H., Li, Q., and Stamatoyannopoulos, G. (1997) Production of transgenic mice with yeast artificial chromosomes. Trends Genet. 13, 61ā66.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 2003 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Damert, A., Kusserow, H. (2003). Generation of Transgenic Mice by Pronuclear Injection. In: Nag, S. (eds) The Blood-Brain Barrier. Methods in Molecular Medicineā¢, vol 89. Humana Press. https://doi.org/10.1385/1-59259-419-0:513
Download citation
DOI: https://doi.org/10.1385/1-59259-419-0:513
Publisher Name: Humana Press
Print ISBN: 978-1-58829-073-1
Online ISBN: 978-1-59259-419-1
eBook Packages: Springer Protocols