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Development of a cellular model to study alternative splicing of type II collagen gene.
J Orthop Res. 2002 May; 20(3):516-9.JO

Abstract

To study regulation of alternative splicing of type II collagen (COL2) pre-mRNA, we constructed a mouse COL2 "minigene" containing genomic sequences spanning exon 1 to exon 4 of COL2 downstream of a cytomegalovirus (CMV) promoter. This minigene was introduced into ATDC5 cells, which undergo chondrocytic differentiation when treated with insulin. Alternative splicing of the COL2 minigene was evaluated by comparing the expression of the two mRNAs transcribed from the minigene to the expression of alternatively spliced transcripts from the endogenous COL2 gene. This analysis suggested that regulation of alternative splicing of pre-mRNAs from the minigene and the endogenous COL2 gene are accomplished by similar mechanisms. We conclude that the cloned genomic fragment contains key sequences necessary for alternative splicing of COL2 pre-mRNA. This system provides a useful model to begin the process of identifying cis- and trans-acting factors that carry out alternative splicing of COL2 pre-mRNA during chondrocyte differentiation.

Authors+Show Affiliations

Department of Orthopedic Research, Mayo Clinic and Foundation, Rochester, MN 55905, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

12038625

Citation

Hatano, H, et al. "Development of a Cellular Model to Study Alternative Splicing of Type II Collagen Gene." Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society, vol. 20, no. 3, 2002, pp. 516-9.
Hatano H, Sarkar G, Bolander ME. Development of a cellular model to study alternative splicing of type II collagen gene. J Orthop Res. 2002;20(3):516-9.
Hatano, H., Sarkar, G., & Bolander, M. E. (2002). Development of a cellular model to study alternative splicing of type II collagen gene. Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society, 20(3), 516-9.
Hatano H, Sarkar G, Bolander ME. Development of a Cellular Model to Study Alternative Splicing of Type II Collagen Gene. J Orthop Res. 2002;20(3):516-9. PubMed PMID: 12038625.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a cellular model to study alternative splicing of type II collagen gene. AU - Hatano,H, AU - Sarkar,G, AU - Bolander,M E, PY - 2002/6/1/pubmed PY - 2002/6/18/medline PY - 2002/6/1/entrez SP - 516 EP - 9 JF - Journal of orthopaedic research : official publication of the Orthopaedic Research Society JO - J Orthop Res VL - 20 IS - 3 N2 - To study regulation of alternative splicing of type II collagen (COL2) pre-mRNA, we constructed a mouse COL2 "minigene" containing genomic sequences spanning exon 1 to exon 4 of COL2 downstream of a cytomegalovirus (CMV) promoter. This minigene was introduced into ATDC5 cells, which undergo chondrocytic differentiation when treated with insulin. Alternative splicing of the COL2 minigene was evaluated by comparing the expression of the two mRNAs transcribed from the minigene to the expression of alternatively spliced transcripts from the endogenous COL2 gene. This analysis suggested that regulation of alternative splicing of pre-mRNAs from the minigene and the endogenous COL2 gene are accomplished by similar mechanisms. We conclude that the cloned genomic fragment contains key sequences necessary for alternative splicing of COL2 pre-mRNA. This system provides a useful model to begin the process of identifying cis- and trans-acting factors that carry out alternative splicing of COL2 pre-mRNA during chondrocyte differentiation. SN - 0736-0266 UR - https://www.unboundmedicine.com/medline/citation/12038625/Development_of_a_cellular_model_to_study_alternative_splicing_of_type_II_collagen_gene_ L2 - https://doi.org/10.1016/S0736-0266(01)00123-1 DB - PRIME DP - Unbound Medicine ER -