Human Molecular Genetics, Vol 7, 2079-2088, Copyright © 1998 by Oxford University Press
KR Peterson, PA Navas, Q Li and G Stamatoyannopoulos
Yeast artificial chromosome (YAC) transgenesis is associated with a high
frequency of deletions in the integrated transgenes. To determine the
impact of these rearrangements on the ability to derive structure- function
relationships using YACs, transgenic mice were generated with 248 or 155 kb
beta-globin locus YACs. The transgenics were examined for structural
integrity of the YAC using an approach of structural analysis that
unambiguously demonstrates intactness of YAC transgene copies. Globin gene
expression per copy of each integrated transgene and the profiles of globin
gene expression during development were determined. Diverse deletion
patterns were observed in one or more integrated YACs in all the 248 and
most of the 155 kb transgenic lines we analyzed. However, when the
structure of the major regulatory element of the beta-globin locus, the
locus control region, was preserved, the genes of the beta-globin locus
functioned normally and globin transgenes of both the 248 and 155 kb
beta-YACs were expressed in a position-independent, copy number-dependent
manner. Furthermore, the globin genes of both beta-YACs displayed normal
developmental regulation. We conclude that YACs can be used for analysis of
structure- function relationships of large genes or multigene loci in spite
of the tendency for rearrangements and deletions of the integrated
transgenes. However, detailed structural evidence for integrity and
continuity of locus sequences is required for correct interpretation of
functional data.
ARTICLES
LCR-dependent gene expression in beta-globin YAC transgenics: detailed structural studies validate functional analysis even in the presence of fragmented YACs
Division of Medical Genetics, Department of Medicine and Department of Genetics, University of Washington, Seattle, WA 98195, USA. kpeterson@kumc.edu
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