Muscle-specific locus control region activity associated with the human desmin gene.
We describe the reproduction of the full pattern of expression of the muscle-specific desmin gene in transgenic mice using a 240-kb genomic clone spanning the human desmin locus. Analysis of RNA from adult tissues demonstrated that this fragment possesses all the necessary genetic regulatory elements required to provide reproducible, site-of-integration-independent, physiological levels of tissue-specific expression that is directly proportional to transgene copy number in all muscle cell types. In situ hybridization revealed that in marked contrast to murine desmin which is strongly expressed in the myotome of the somites, skeletal muscles, the heart, and smooth muscle of the vasculature by 9.5 days postcoitum, human desmin transgene expression was completely absent from smooth muscles, was very weak and restricted to the atrium and outflow tract within the heart, and was expressed at only 5% of murine desmin mRNA levels within the myotome of the somites. The spatial distribution and levels of human and mouse desmin expression were not coincident until 14.5 days postcoitum. Immunohistochemical analysis of human embryos at comparable stages of development showed that this transgene faithfully reproduces the human and not the mouse developmental expression pattern for this gene in transgenic mice. These results indicate that the 240-kb desmin genomic clone is capable of establishing an independent, chromatin domain in transgenic mice and provides the first definitive data for muscle-specific locus control region activity. In addition, our results demonstrate that the behavior of human transgenes in mice should, whenever possible, be compared to expression patterns for that gene in human embryonic as well as adult tissues.
Raguz
S
S
UMDS, Guy's Hospital, London Bridge, London, SE1 9RT, United Kingdom.
Hobbs
C
C
Yagüe
E
E
Ioannou
P A
PA
Walsh
F S
FS
Antoniou
M
M
D000818:Animals; D003001:Cloning, Molecular; D003893:Desmin / Q000096:biosynthesis / Q000235:genetics*; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005314:Embryonic and Fetal Development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D015698:Genomic Library; D006579:Heterozygote; D006801:Humans; D017403:In Situ Hybridization; D019901:Locus Control Region; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D024510:Muscle Development; D009132:Muscles / Q000196:embryology / Q000378:metabolism*; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping
D003893:Desmin; D011994:Recombinant Proteins