Taking control of gene expression with light-activated oligonucleotides
Taking control of gene expression with light-activated oligonucleotides
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DOI：
10.2144/000112519
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发表时间：
2007-08-01
期刊：
影响因子：
2.7
通讯作者：
Tang, XinJing
中科院分区：
文献类型：
--
作者：
Dmochowski, Ivan J.;Tang, XinJing
被紫外光有效激活的笼状寡核苷酸的最新发展为以非常高的空间和时间分辨率调控基因表达创造了机会。通过选择性地调节基因活性，这些光化学工具将促进阐明基因功能的努力，并最终可能服务于治疗目的。我们演示了在DNA双链中加入可光切割的封闭基团如何瞬间阻止DNA聚合酶的活性。事实上，笼子里的寡核苷酸使控制许多不同的蛋白质-寡核苷酸相互作用成为可能。在相关实验中，将反向互补(反义)寡核苷酸与靶mRNA杂交，可以通过招募包涵体核糖核酸酶或立体阻断核糖体来抑制翻译。本实验室最近合成了由硫代磷酸或肽核酸(PNA)组成的笼状反义寡核苷酸。反链寡核苷酸通过光切连接子连接到互补的阻断寡核链上，不能与靶基因结合。这为反义基因的光调控杂交提供了一种有用的方法。TRANCL为靶向的mRNA。笼式DNA和PNA寡核苷酸已被证明在细胞和斑马鱼胚胎中光调控基因表达方面有效。
The recent development of caged oligonucleotides that are, efficiently activated by ultraviolet (UV) light creates opportunities for regulating gene expression with very high spatial and temporal resolution. By selectively modulating gene activity, these photochemical tools will facilitate efforts to elucidate gene function and may eventually serve therapeutic aims. We demonstrate how the incorporation of a photocleavable blocking group within a DNA duplex can transiently arrest DNA polymerase activity. Indeed, caged oligonucleotides make it possible to control many different protein-oligonucleotide interactions. In related experiments hybridization of a reverse complimentary (antisense) oligodeoxynucleotide to target mRNA can inhibit translation by recruiting enclogenous RNases or sterically blocking the ribosome. Our laboratory recently synthesized caged antisense oligonucleoticles composed of phosphorothioated DNA or pepticle nucleic acid (PNA). The antiserise oligonucleoticle, which was attached to i a complementary blocking oligonuclecticle strand by a photocleavable linker, was blocked from binding target mRNA. This provided a useful method for photomodulating hybridization of the antisense. trancl to target mRNA. Caged DNA and PNA oligonucleoticles have proven effective at photoregulating gene expression in cells and zebrafish embryos.