Nucleic Acids Research, Vol 26, Issue 12 2849-2858, Copyright © 1998 by Oxford University Press
TA Graubert, BA Hug, R Wesselschmidt, CL Hsieh, TM Ryan, TM Townes and TJ Ley
The random insertion of transgenes into the genomic DNA of mice usually
leads to widely variable levels of expression in individual founder lines.
To study the mechanisms that cause variegation, we designed a transgene
that we expected to variegate, which consisted of a beta- globin locus
control region 5' HS-2 linked in tandem to a tagged human beta-globin gene
(into which a Lac-Z cassette had been inserted). All tested founder lines
exhibited red blood cell-specific expression, but levels of expression
varied >1000-fold from the lowest to the highest expressing line. Most
of the variation in levels of expression appeared to reflect differences in
the percentage of cells in the peripheral blood that expressed the
transgene, which ranged from 0.3% in the lowest expressing line to 88% in
the highest; the level of transgene expression per cell varied no more than
10-fold from the lowest to the highest expressing line. These differences
in expression levels could not be explained by the location of transgene
integration, by an effect of beta-galactosidase on red blood cell survival,
by the half life of the beta-galactosidase enzyme or by the age of the
animals. The progeny of all early erythroid progenitors (BFU-E
colony-forming cells) exhibited the same propensity to variegate in
methylcellulose-based cultures, suggesting that the decision to variegate
occurs after the BFU-E stage of erythroid differentiation. Collectively,
these data suggest that variegation in levels of transgene expression are
due to local, integration site-dependent phenomena that alter the
probability that a transgene will be expressed in an appropriate cell;
however, these local effects have a minimal impact on the transgene's
activity in the cells that initiate transcription.
ARTICLES
Stochastic, stage-specific mechanisms account for the variegation of a human globin transgene
Division of Bone Marrow Transplantation and Stem Cell Biology, Departments of Medicine and Genetics, Washington University School of Medicine, Campus Box 8007, 660 South Euclid Avenue, St Louis, MO 63110- 1093, USA. timley@im.wustl.edu
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