In vivo AGO-APP identifies a module of microRNAs cooperatively controlling exit from neural stem cell state

Author:

Narbonne-Reveau KarineORCID,Erni AndreaORCID,Eichner Norbert,Surbhi Surbhi,Meister Gunter,Beclin ChristopheORCID,Maurange CédricORCID,Cremer HaroldORCID

Abstract

AbstractMicroRNAs (miRNAs) are essential regulators of all developmental processes. Their function is particularly important during neurogenesis, when the production of large numbers of neurons from a limited number of neural stem cells depends on the precise control of determination, proliferation and differentiation. However, miRNA regulation of target mRNAs is highly promiscuous, one miRNA can target many mRNAs and vice versa, raising the question of how specificity is achieved to elicit a precise regulatory response.Here we introduce AGO-APP, a novel approach to purify Argonaute-bound miRNAs directly from cells and tissues in vivo, to isolate actively inhibiting miRNAs from different neural cell populations in the larval Drosophila central nervous system. We identify a defined group of miRNAs that redundantly target all iconic genes known to control the transition from neuroblasts to neurons. In vivo functional studies demonstrate that knockdown of individual miRNAs does not induce detectable cellular phenotypes. However, simultaneous knockdown of multiple miRNAs leads to precocious stem cell differentiation, demonstrating functional interdependence. Thus, miRNAs cooperate within a regulatory module to specify the targeted gene network.

Publisher

Cold Spring Harbor Laboratory

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