Dynamic Nanostructures for Conditional Activation and Deactivation of Biological Pathways.
Dynamic Nanostructures for Conditional Activation and Deactivation of Biological Pathways.
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用于生物途径的条件激活和失活的动态纳米结构。
DOI：
10.1007/978-1-0716-3417-2_22
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Afonin,KirillA
中科院分区：
文献类型：
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作者：
Radwan,Yasmine;Rebolledo,LauraP;Panigaj,Martin;Afonin,KirillA
核酸纳米技术利用天然和合成结构基序来构建多功能核酸纳米颗粒（NANPs）。这些合理设计的组件可以进一步配备功能性核酸和其他分子，如肽、荧光染料等。除了直接与调控的靶基因转录物相互作用的核酸外，NANPs还可以显示诱饵，其中寡核苷酸与转录因子结合序列延伸，阻止转录起始。活化B细胞的核因子kappa-轻链增强子（NF-κB）是调节炎性疾病和癌症发病机制的五种关键转录因子；因此，它们是治疗的相关目标。一种治疗方法涉及相互依赖的自我识别杂交DNA/RNA纤维，旨在结合NF-κB并阻止其与炎症反应中NF-κB依赖性基因的启动子区域相互作用。诱骗NF-κB导致无法启动受调控基因的转录，这是一种很有前途的基因调控和基因治疗方法。本文描述的方案提供了合成NF-κB诱饵纤维的详细步骤，以及使用聚丙烯酰胺凝胶电泳（以确认所需的物理化学性质和纯度）和功能生物测定（以确认所需的生物活性）对其进行表征。
Nucleic acid nanotechnology utilizes natural and synthetic structural motifs to build versatile nucleic acid nanoparticles (NANPs). These rationally designed assemblies can be further equipped with functional nucleic acids and other molecules such as peptides, fluorescent dyes, etc. In addition to nucleic acids that directly interact with the regulated target gene transcripts, NANPs can display decoys, wherein the oligonucleotide stretches with transcription factor binding sequences, preventing transcription initiation. The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a group of five crucial transcription factors regulating the pathogenesis of inflammatory diseases and cancer; as such, they are relevant targets for therapy. One therapeutic approach involves interdependent self-recognizing hybridized DNA/RNA fibers designed to bind NF-κB and prevent its interaction with the promotor region of NF-κB-dependent genes involved in inflammatory responses. Decoying NF-κB results in the inability to initiate transcription of regulated genes, showing a promising approach to gene regulation and gene therapy. The protocol described herein provides detailed steps for the synthesis of NF-κB decoy fibers, as well as their characterization using polyacrylamide gel electrophoresis (to confirm desired physicochemical properties and purity) and functional bioassays (to confirm desired biological activity).