Upstream regulatory elements necessary for expression of the rat COL1A1 promoter in transgenic mice

Zoran Bogdanovic, Antonio Bedalov, Paul H. Krebsbach, Dubravko Pavlin, Charles O. Woody, Stephen H. Clark, Huw F. Thomas, David W. Rowe, Barbara E. Kream, Alexander C. Lichtler

Research output: Contribution to journalArticle

89 Scopus citations

Abstract

The activity of fusion genes containing fragments of the COL1A1 promoter was measured in tissues from 6‐ to 8‐day‐old transgenic mice. ColCAT3.6 contains approximately 3.6 kb (‐3521 to 115 bp) of the rat COL1A1 gene, the chloramphenicol acetyltransferase (CAT) reporter gene, and the SV40 splice and polyadenylation sequences. ColCAT2.3 and ColCAT1.7 are deletion constructs that contain 2296 and 1667 bp of COL1A1 upstream from the RNA start site, respectively. For each transgene, up to six lines of mice were characterized. Both ColCAT3.6 and ColCAT2.3 had similar activity in bone and tooth; ColCAT1.7 was inactive. In transgenic calvariae, levels of transgene mRNA paralleled levels of CAT activity. In tendon, the activity of ColCAT2.3 was 3‐ to 4‐fold lower than that of ColCAT3.6, and the activity ColCAT1.7 was 16‐fold lower than that of ColCAT2.3. There was little activity of the ColCAT constructs in liver and brain. These data show that DNA sequnces between −2.3 and −1.7 kb are required for COL1A1 promoter expression in bone and tooth; sequences that control expression in tendon are distributed between −3.5 and −1.7 kb of the promoter, with sequences downstream of −1.7 kb still capable of directing expression to this tissue. The cis elements that govern basal expression of COL1A1 in transgenic calvariae appear to be different from those required for optimal expression of the COL1A1 promoter in stably transfected osteoblastic cells.

Original languageEnglish (US)
Pages (from-to)285-292
Number of pages8
JournalJournal of Bone and Mineral Research
Volume9
Issue number2
DOIs
StatePublished - Feb 1994

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

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    Bogdanovic, Z., Bedalov, A., Krebsbach, P. H., Pavlin, D., Woody, C. O., Clark, S. H., Thomas, H. F., Rowe, D. W., Kream, B. E., & Lichtler, A. C. (1994). Upstream regulatory elements necessary for expression of the rat COL1A1 promoter in transgenic mice. Journal of Bone and Mineral Research, 9(2), 285-292. https://doi.org/10.1002/jbmr.5650090218