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A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice.
J Cell Sci. 1995 Dec; 108 (Pt 12):3677-84.JC

Abstract

Type II collagen is a major chondrocyte-specific component of the cartilage extracellular matrix and it represents a typical differentiation marker of mature chondrocytes. In order to delineate cis-acting elements of the mouse pro alpha 1(II) collagen gene that control chondrocyte-specific expression in intact mouse embryos, we generated transgenic mice harboring chimeric constructions in which varying lengths of the promoter and intron 1 sequences were linked to a beta-galactosidase reporter gene. A construction containing a 3,000 bp promoter and a 3,020 bp intron 1 fragment directed high levels of beta-galactosidase expression specifically to chondrocytes. Expression of the transgene coincided with the temporal expression of the endogenous gene at all stages of embryonic development. Successive deletions of intron 1 delineated a 182 bp fragment which targeted beta-galactosidase expression to chondrocytes with the same specificity as the larger intron 1 fragment. Transgenic mice harboring a 309 bp Col2a1 promoter lacking intron 1 tester sequences showed no beta-galactosidase expression in chondrocytes. Reduction of the 182 bp fragment to a 73 bp subfragment surrounding a decamer sequence previously reported to be involved in chondrocyte specificity, resulted in loss of transgene expression in chondrocytes. When the Col2a1 promoter was replaced with a minimal beta-globin promoter, the 182 bp intron 1 sequence was still able to target expression of the transgene to chondrocytes. We conclude that a 182 bp intron 1 DNA segment of the mouse Col2a1 gene contains the necessary information to confer high-level, temporally correct, chondrocyte expression on a reporter gene in intact mouse embryos and that Col2a1 promoter sequences are dispensable for chondrocyte expression.

Authors+Show Affiliations

Department of Molecular Genetics, University of Texas, M. D. Anderson Cancer Center, Houston 77030, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8719874

Citation

Zhou, G, et al. "A 182 Bp Fragment of the Mouse Pro Alpha 1(II) Collagen Gene Is Sufficient to Direct Chondrocyte Expression in Transgenic Mice." Journal of Cell Science, vol. 108 (Pt 12), 1995, pp. 3677-84.
Zhou G, Garofalo S, Mukhopadhyay K, et al. A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice. J Cell Sci. 1995;108 (Pt 12):3677-84.
Zhou, G., Garofalo, S., Mukhopadhyay, K., Lefebvre, V., Smith, C. N., Eberspaecher, H., & de Crombrugghe, B. (1995). A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice. Journal of Cell Science, 108 (Pt 12), 3677-84.
Zhou G, et al. A 182 Bp Fragment of the Mouse Pro Alpha 1(II) Collagen Gene Is Sufficient to Direct Chondrocyte Expression in Transgenic Mice. J Cell Sci. 1995;108 (Pt 12):3677-84. PubMed PMID: 8719874.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice. AU - Zhou,G, AU - Garofalo,S, AU - Mukhopadhyay,K, AU - Lefebvre,V, AU - Smith,C N, AU - Eberspaecher,H, AU - de Crombrugghe,B, PY - 1995/12/1/pubmed PY - 1995/12/1/medline PY - 1995/12/1/entrez SP - 3677 EP - 84 JF - Journal of cell science JO - J Cell Sci VL - 108 (Pt 12) N2 - Type II collagen is a major chondrocyte-specific component of the cartilage extracellular matrix and it represents a typical differentiation marker of mature chondrocytes. In order to delineate cis-acting elements of the mouse pro alpha 1(II) collagen gene that control chondrocyte-specific expression in intact mouse embryos, we generated transgenic mice harboring chimeric constructions in which varying lengths of the promoter and intron 1 sequences were linked to a beta-galactosidase reporter gene. A construction containing a 3,000 bp promoter and a 3,020 bp intron 1 fragment directed high levels of beta-galactosidase expression specifically to chondrocytes. Expression of the transgene coincided with the temporal expression of the endogenous gene at all stages of embryonic development. Successive deletions of intron 1 delineated a 182 bp fragment which targeted beta-galactosidase expression to chondrocytes with the same specificity as the larger intron 1 fragment. Transgenic mice harboring a 309 bp Col2a1 promoter lacking intron 1 tester sequences showed no beta-galactosidase expression in chondrocytes. Reduction of the 182 bp fragment to a 73 bp subfragment surrounding a decamer sequence previously reported to be involved in chondrocyte specificity, resulted in loss of transgene expression in chondrocytes. When the Col2a1 promoter was replaced with a minimal beta-globin promoter, the 182 bp intron 1 sequence was still able to target expression of the transgene to chondrocytes. We conclude that a 182 bp intron 1 DNA segment of the mouse Col2a1 gene contains the necessary information to confer high-level, temporally correct, chondrocyte expression on a reporter gene in intact mouse embryos and that Col2a1 promoter sequences are dispensable for chondrocyte expression. SN - 0021-9533 UR - https://www.unboundmedicine.com/medline/citation/8719874/A_182_bp_fragment_of_the_mouse_pro_alpha_1_II__collagen_gene_is_sufficient_to_direct_chondrocyte_expression_in_transgenic_mice_ L2 - http://jcs.biologists.org/cgi/pmidlookup?view=long&pmid=8719874 DB - PRIME DP - Unbound Medicine ER -