Centro Andaluz de Biologia del Desarrollo, CSIC-Universidad Pablo de Olavide-Junta de Andalucía, Sevilla, Spain.
PLoS One. 2011;6(9):e24824. doi: 10.1371/journal.pone.0024824. Epub 2011 Sep 13.
Non-coding DNA conservation across species has been often used as a predictor for transcriptional enhancer activity. However, only a few systematic analyses of the function of these highly conserved non-coding regions (HCNRs) have been performed. Here we use zebrafish transgenic assays to perform a systematic study of 113 HCNRs from human chromosome 16. By comparing transient and stable transgenesis, we show that the first method is highly inefficient, leading to 40% of false positives and 20% of false negatives. When analyzed in stable transgenic lines, a great majority of HCNRs were active in the central nervous system, although some of them drove expression in other organs such as the eye and the excretory system. Finally, by testing a fraction of the HCNRs lacking enhancer activity for in vivo insulator activity, we find that 20% of them may contain enhancer-blocking function. Altogether our data indicate that HCNRs may contain different types of cis-regulatory activity, including enhancer, insulators as well as other not yet discovered functions.
跨物种的非编码 DNA 保守性经常被用作转录增强子活性的预测因子。然而，只有少数对这些高度保守的非编码区域（HCNRs）的功能进行了系统分析。在这里，我们使用斑马鱼转基因实验来对来自人类 16 号染色体的 113 个 HCNRs 进行系统研究。通过比较瞬时和稳定的转基因，我们表明第一种方法效率非常低，导致 40%的假阳性和 20%的假阴性。当在稳定的转基因系中进行分析时，绝大多数 HCNRs 在中枢神经系统中具有活性，尽管其中一些在眼睛和排泄系统等其他器官中驱动表达。最后，通过测试缺乏增强子活性的 HCNRs 中一部分的体内绝缘子活性，我们发现其中 20%可能含有增强子阻断功能。总的来说，我们的数据表明 HCNRs 可能包含不同类型的顺式调控活性，包括增强子、绝缘子以及其他尚未发现的功能。
