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© The Rockefeller University Press, 0021-9525/1998//1291 $5.00
The Journal of Cell Biology, Volume 141, Number 6, , 1998 1291-1300

Articles

Different cis-Regulatory DNA Elements Mediate Developmental Stage- and Tissue-specific Expression of the Human COL2A1 Gene in Transgenic Mice



Keith K.H. Leung*, Ling Jim Ng*, Ken K.Y. Ho*, Patrick P.L. Tam{ddagger}, and Kathryn S.E. Cheah*

* Department of Biochemistry, The University of Hong Kong, Hong Kong; and {ddagger} Embryology Unit, Children's Medical Research Institute, New South Wales 2145, Australia

Expression of the type II collagen gene (human COL2A1, mouse Col2a1) heralds the differentiation of chondrocytes. It is also expressed in progenitor cells of some nonchondrogenic tissues during embryogenesis. DNA sequences in the 5' flanking region and intron 1 are known to control tissue-specific expression in vitro, but the regulation of COL2A1 expression in vivo is not clearly understood. We have tested the regulatory activity of DNA sequences from COL2A1 on the expression of a lacZ reporter gene in transgenic mice. We have found that type II collagen characteristic expression of the transgene requires the enhancer activity of a 309-bp fragment (+2,388 to +2,696) in intron 1 in conjunction with 6.1-kb 5' sequences. Different regulatory elements were found in the 1.6-kb region (+701 to +2,387) of intron 1 which only needs 90-bp 5' sequences for tissue-specific expression in different components of the developing cartilaginous skeleton. Distinct positive and negative regulatory elements act together to control tissue-specific transgene expression in the developing midbrain neuroepithelium. Positive elements affecting expression in the midbrain were found in the region from –90 to –1,500 and from +701 to +2,387, whereas negatively acting elements were detected in the regions from –1,500 to –6,100 and +2,388 to +2,855.

Abbreviations used in this paper: dpc, day(s) post coitum; nt, nucleotide(s).

Address all correspondence to Professor Kathryn S.E. Cheah, Department of Biochemistry, The University of Hong Kong, Sassoon Road, Hong Kong. Tel.: (852) 2819-9240 or 2819-9170. Fax: (852) 2855-1254. E-mail: hrmbdkc{at}hkumd1.hku.hk


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