Different cis-regulatory DNA elements mediate developmental stage- and tissue-specific expression of the human COL2A1 gene in transgenic mice.
Different cis-regulatory DNA elements mediate developmental stage- and tissue-specific expression of the human COL2A1 gene in transgenic mice.
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不同的顺式调节DNA元件介导了转基因小鼠中人Col2A1基因的发育阶段和组织特异性表达。
DOI：
10.1083/jcb.141.6.1291
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发表时间：
1998-06-15
期刊：
影响因子：
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通讯作者：
Cheah KS
中科院分区：
文献类型：
--
作者：
Leung KK;Ng LJ;Ho KK;Tam PP;Cheah KS
II型胶原基因（人Col 2A 1，小鼠Col 2a 1）的表达预示着软骨细胞的分化。它还在胚胎发生期间的一些非软骨形成组织的祖细胞中表达。已知5′侧翼区和内含子1中的DNA序列在体外控制组织特异性表达，但对体内COL 2A 1表达的调控尚不清楚。我们已经测试了来自COL 2A 1的DNA序列对转基因小鼠中lacZ报告基因表达的调节活性。我们发现，II型胶原蛋白的特征性表达需要内含子1中的309 bp片段（+2，388至+2，696）与6.1 kb 5′序列结合的增强子活性。在内含子1的1.6kb区域（+701至+2，387）发现了不同的调控元件，其仅需要90 bp的5′序列就可以在发育中的软骨骨骼的不同组分中组织特异性表达。不同的正调控元件和负调控元件共同作用以控制发育中的中脑神经上皮中的组织特异性转基因表达。影响中脑表达的正向元素出现在-90到-1,500和+701到+2,387区域，而负向元素出现在-1,500到-6,100和+2,388到+2855区域。
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.