Lack of a phenotype in transgenic mice aberrantly expressing COL2A1 mRNA because of highly selective post-transcriptional down-regulation.
We reported previously that a 1.9-kb 5'-fragment from the human COL1A1 gene drove transcription of a promoterless human COL2A1 gene in tissues of transgenic mice that normally express the COL1A1 but not the COL2A1 gene. In the present study, we have established that the aberrant transcription of the COL2A1 gene did not produce any gross or microscopic phenotype, because the transcripts were not efficiently translated in cells that do not normally express the COL2A1 gene. In two lines of transgenic mice, the mRNA levels from the transgene were 30% to 45% of the mRNA for the proalpha1(I) chain of type I procollagen, the most abundant mRNA in the same tissues. Analysis of collagens extracted from skin of the transgenic mice indicated that triple-helical type II collagen, with the normal pattern of cyanogen bromide peptides, was synthesized from the transgene. However, the level of type II collagen in skin was less than 2% of the level of type I collagen. Hybridization in situ indicated the presence of mRNA for both COL2A1 and COL1A1 in the same cells. Immunofluorescence staining for type II collagen, however, was negative in the same tissues. The results, therefore, indicated that many mesenchymal cells in the transgenic mice had high steady-state levels of the homologous mRNAs for type I and type II procollagen, but only the mRNAs for type I procollagen were efficiently translated.
Yuan
C M
CM
Center for Gene Therapy, MCP Hahnemann University, Philadelphia, PA 19102, USA.
Ala-Kokko
L
L
Le Guellec
D
D
Franc
S
S
Fertala
A
A
Khillan
J S
JS
Sokolov
B P
BP
Prockop
D J
DJ
D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics* / Q000648:ultrastructure; D015536:Down-Regulation; D005347:Fibroblasts / Q000166:cytology; D005455:Fluorescent Antibody Technique; D017403:In Situ Hybridization; D008334:Mandible / Q000648:ultrastructure; D051379:Mice; D008822:Mice, Transgenic; D010641:Phenotype; D011347:Procollagen / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012867:Skin / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D013710:Tendons / Q000648:ultrastructure; D014070:Tooth / Q000648:ultrastructure; D014989:Xiphoid Bone / Q000648:ultrastructure
D011347:Procollagen; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D003094:Collagen