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Production of YAC transgenic mice by pronuclear injection

Ainscough, Justin, John, Rosalind Margaret ORCID: and Barton, Sheila C. 2002. Production of YAC transgenic mice by pronuclear injection. Ward, Andrew, ed. Genomic Imprinting, Vol. 181. Methods in Molecular Biology, Humana Press, pp. 55-65. (10.1385/1-59259-211-2:55)

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The production of transgenic mice using small DNA constructs has been widely used for many years to investigate the regulation of gene activity. Small plasmid-based constructs (less than 20 kb) have been favored for a number of reasons, particularly the ease with which they can be manipulated and purified in large quantities. While this approach is powerful, there are some problems associated with the size of these transgenes. In particular, many of these small transgenes do not reproduce accurately the expression seen from the endogenous gene. For some genes the regulatory elements that control activity are located at a distance from the promoter and can be omitted from the transgene. These may be enhancers, repressors, boundary elements, or even locus control regions (LCRs), which are responsible for maintaining the correct spatial and temporal expression patterns of a number of genes, such as the globin clusters in mouse and humans (1). More important, small transgenes are susceptible to position effects from the chromatin environment in which they integrate, which often results in either ectopic expression (from trapping of nearby enhancers for other genes) or suppression of gene activity. Finally, small transgenes usually integrate in a multicopy tandem arrangement that does not accurately reflect the situation seen at the endogenous locus.

Item Type: Book Section
Date Type: Publication
Status: Published
Schools: Biosciences
Subjects: Q Science > QH Natural history > QH426 Genetics
Publisher: Humana Press
ISBN: 9780896037410
ISSN: 1064-3745
Last Modified: 28 Oct 2022 08:33

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