Photocaged DNAzymes as a general method for sensing metal ions in living cells.
Photocaged DNAzymes as a general method for sensing metal ions in living cells.
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DOI：
10.1002/anie.201408333
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发表时间：
2014-12-08
影响因子：
16.6
通讯作者：
Lu, Yi
中科院分区：
文献类型：
--
作者：
Hwang, Kevin;Wu, Peiwen;Kim, Taejin;Lei, Lei;Tian, Shiliang;Wang, Yingxiao;Lu, Yi
DNA酶是具有催化活性的DNA序列，已被证明是一种潜在的检测多种金属离子的平台。尽管有很大的希望，但使用DNA酶的细胞传感一直很困难，主要是由于第一代DNA酶传感器的“永远在线”性质。为了克服这一限制，我们在本文中展示的设计和合成的光活化或photocaged的DNA酶，其应用在传感Zn（II）在活细胞中。在这种设计中，8-17 DNA酶易断裂位置的腺苷核糖核苷酸被2′-O-硝基苄基腺苷取代，使DNA酶失活，从而使其完整地递送到细胞中，防止细胞内的非特异性降解。在365 nm处的照射恢复DNAzyme活性，允许对DNAzyme对金属离子的感测活性进行时间控制。同样的策略也被应用于GR-5 DNA酶检测铅（II），证明其广泛的普适性。
DNAzymes, sequences of DNA with catalytic activity, have been demonstrated as a potential platform for sensing a wide range of metal ions. Despite significant promise, cellular sensing using DNAzymes has been difficult, mainly due to the ‘always-on’ nature of first generation DNAzyme sensors. To overcome this limitation, we demonstrate herein the design and synthesis of a photoactivatable or photocaged DNAzyme, and its application in sensing Zn(II) in living cells. In this design the adenosine ribonucleotide at the scissile position of the 8–17 DNAzyme is replaced by 2′-O-nitrobenzyl adenosine, rendering the DNAzyme inactive and thus allowing its delivery into cells intact, protected from non-specific degradation within cells. Irradiation at 365 nm restores DNAzyme activity, allowing for temporal control over the DNAzyme’s sensing activity for metal ions. The same strategy has also been applied towards the GR-5 DNAzyme for detection of Pb(II), demonstrating its broad generalizability.