Handcuffed antisense oligonucleotides for light-controlled cell-free expression.
Handcuffed antisense oligonucleotides for light-controlled cell-free expression.
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用于光控无细胞表达的手铐反义寡核苷酸。
DOI：
10.1039/d3cc01374j
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Hartmann D
中科院分区：
文献类型：
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作者：
Hartmann D
开发利用光笼沉默反义寡核苷酸(ASO)的简单方法，为精确调控生物系统打开了可能性。在这里，我们开发了一种基于将两个ASO‘铐’到一个蛋白质上的光笼策略。沉默是通过两个末端可光切割的生物素修饰的ASO与一个链霉亲和素的二价结合实现的。这些“戴着手铐”的寡核苷酸显示，在无细胞蛋白质合成中，基因敲除活性急剧下降，并通过光照解锁，重新获得全部活性。
Developing simple methods to silence antisense oligonucleotides (ASOs) using photocages opens up the possibility of precise regulation of biological systems. Here, we have developed a photocaging strategy based on ‘handcuffing’ two ASOs to a protein. Silencing was achieved by divalent binding of two terminally photocleavable biotin-modified ASOs to a single streptavidin. These ‘handcuffed’ oligonucleotides showed a drastic reduction in gene knockdown activity in cell-free protein synthesis and were unlocked through illumination, regaining full activity.