An oligonucleotide microarray for microRNA expression analysis based on labeling RNA with quantum dot and nanogold probe.
An oligonucleotide microarray for microRNA expression analysis based on labeling RNA with quantum dot and nanogold probe.
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基于量子点和纳米金探针标记 RNA 的用于 microRNA 表达分析的寡核苷酸微阵列。
DOI：
10.1093/nar/gni019
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发表时间：
2005-01-31
影响因子：
14.9
通讯作者：
Ruan KC
中科院分区：
文献类型：
--
作者：
Liang RQ;Li W;Li Y;Tan CY;Li JX;Jin YX;Ruan KC
微小RNA（microRNAs，miRNAs）通过靶向mRNA进行切割或翻译抑制，在动物和植物中发挥重要的调控作用。它们具有不同的表达模式，可能调节多种发育和生理过程。分析miRNA的表达谱有助于研究miRNA的生物学功能。我们报道了一种新的miRNA谱芯片，该芯片中miRNA的3′端直接用生物素标记，并与固定在载玻片上的互补寡核苷酸探针杂交，随后通过链霉亲和素标记的量子点与miRNA通过链霉亲和素-生物素相互作用结合来检测荧光。该微阵列对miRNA的检测限为0.4fmol，检测动态范围约为2个数量级。我们构建了一个基因芯片模型，对水稻叶片和根中的11个miRNAs进行了分析。SSP. indica）幼苗。miRNAs的分析结果具有良好的重复性，与北方印迹结果一致。为了避免使用高成本的检测设备，还成功地将基于纳米金探针结合银增强的比色检测方法引入到miRNA谱微阵列检测中。
MicroRNAs (miRNAs) play important regulatory roles in animals and plants by targeting mRNAs for cleavage or translational repression. They have diverse expression patterns and might regulate various developmental and physiological processes. Profiling miRNA expression is very helpful for studying biological functions of miRNAs. We report a novel miRNA profiling microarray, in which miRNAs were directly labeled at the 3′ terminus with biotin and hybridized with complementary oligo-DNA probes immobilized on glass slides, and subsequently detected by measuring fluorescence of quantum dots labeled with streptavidin bound to miRNAs through streptavidin–biotin interaction. The detection limit of this microarray for miRNA was ∼0.4 fmol, and the detection dynamic range spanned about 2 orders of magnitude. We made a model microarray to profile 11 miRNAs from leaf and root of rice (Oryza sativa L. ssp. indica) seedlings. The analysis results of the miRNAs had a good reproducibility and were consistent with the northern blot result. To avoid using high-cost detection equipment, colorimetric detection, a method based on nanogold probe coupled with silver enhancement, was also successfully introduced into miRNA profiling microarray detection.
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Griffiths-Jones, S
通讯作者：
Griffiths-Jones, S
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