ACS Chem Biol. 2011 Dec 16;6(12):1332-8. doi: 10.1021/cb200290e. Epub 2011 Oct 11.
MicroRNAs (miRNAs) are small non-coding RNAs that play numerous important roles in physiology and human diseases. During animal development, many miRNAs are expressed continuously from early embryos throughout adults, yet it is unclear whether these miRNAs are actually required at all the stages of development. Current techniques of manipulating microRNA function lack the required spatial and temporal resolution to adequately address the functionality of a given microRNA at a specific time or at single-cell resolution. To examine stage- or cell-specific function of miRNA during development and to achieve precise control of miRNA activity, we have developed photoactivatable antisense oligonucleotides against miRNAs. These caged oligonucleotides can be activated with 365 nm light with extraordinarily high efficiency to release potent antisense reagents to inhibit miRNAs. Initial application of these caged antimirs in a model organism (C. elegans) revealed that the activity of a miRNA (lsy-6) is required specifically around the comma stage during embryonic development to control a left/right asymmetric differentiation program in the C. elegans nervous system. This suggests that a transient input of lsy-6 during development is sufficient to specify the neuronal cell fate.
MicroRNAs (miRNAs) 是小的非编码 RNA，在生理和人类疾病中发挥着许多重要作用。在动物发育过程中，许多 miRNA 从早期胚胎持续表达到成年，但尚不清楚这些 miRNA 是否在发育的所有阶段都实际需要。目前操纵 miRNA 功能的技术缺乏所需的时空分辨率，无法充分解决特定时间或单细胞分辨率下特定 miRNA 的功能。为了研究 miRNA 在发育过程中的阶段特异性或细胞特异性功能，并实现 miRNA 活性的精确控制，我们开发了针对 miRNA 的光激活反义寡核苷酸。这些被笼蔽的寡核苷酸可以用 365nm 光以极高的效率激活，释放出有效的反义试剂来抑制 miRNA。这些被笼蔽的 antimirs 在模式生物（秀丽隐杆线虫）中的初步应用表明，miRNA（lsy-6）的活性在胚胎发育过程中特定地在逗号阶段需要，以控制秀丽隐杆线虫神经系统中的左右不对称分化程序。这表明发育过程中 lsy-6 的短暂输入足以指定神经元的细胞命运。
