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Methods Mol Biol ; 2106: 151-159, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31889256

RESUMEN

The Argonaute (AGO) family of proteins plays an essential role in the process of microRNA (miRNA)-mediated gene silencing. More specifically, they are the only known proteins to associate directly with miRNAs within the RNA-induced silencing complex (RISC). Given the importance of miRNA regulation of the transcriptome and its vast implications for human disease, it is essential to understand the molecular underpinnings of miRNA-AGO interactions. Although there are methods available to investigate mature miRNA decay and loading onto AGO2, no feasible method exists to detail the opposite process: release of miRNA from associated AGO proteins. In this chapter, we describe in detail a methodology derived from biochemical approaches, which can be used to quantify the release of any given miRNA from AGOs.


Asunto(s)
Proteínas Argonautas/metabolismo , MicroARNs/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Animales , Línea Celular , Humanos , Cinética , Unión Proteica
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