In vitro selection of signaling aptamers
In vitro selection of signaling aptamers
复制标题
DOI：
10.1038/82414
复制
发表时间：
2000-12-01
影响因子：
46.9
通讯作者：
Ellington, AD
中科院分区：
文献类型：
--
作者：
Jhaveri, S;Rajendran, M;Ellington, AD
可以直接将分子识别转换为光信号的无试剂生物传感器应该会促进用于各种分析物的传感器阵列的发展。与配体紧密结合和特异结合的核酸适体可以很容易地选择，但可能很难适应生物传感器的应用。因此，我们试图开发将适配子的广泛分子识别特性与信号转导相结合的选择方法。抗腺苷适配子是从一个倾斜的池中挑选出来的，这个池含有非常少的荧光尿苷。初级适配子家族在同源分析物ATP的饱和浓度下显示出加倍的相对荧光强度，并且可以检测到低至25微米的ATP浓度。在最好的信号适配子中只有一个尿苷。令人惊讶的是，其他染料可以替代荧光素，并仍然特异性地发出ATP存在的信号，这表明单一尿苷起到了将分子识别转换为光学信号的一般“开关”的作用。
Reagentless biosensors that can directly transduce molecular recognition to optical signals should potentiate the development of sensor arrays for a wide variety of analytes. Nucleic acid aptamers that bind ligands tightly and specifically can be readily selected, but may prove difficult to adapt to biosensor applications. We have therefore attempted to develop selection methods that couple the broad molecular recognition properties of aptamers with signal transduction. Anti-adenosine aptamers were selected from a pool that was skewed to contain very few fluoresceinated uridines. The primary family of aptamers showed a doubling of relative fluorescence intensity at saturating concentrations of a cognate analyte, ATP, and could sense ATP concentrations as low as 25 muM. A single uridine was present in the best signaling aptamer. Surprisingly, other dyes could substitute for fluorescein and still specifically signal the presence of ATP, indicating that the single uridine functioned as a general "switch" for transducing molecular recognition to optical signals.