In vivo AGO-APP for cell-type- and compartment-specific miRNA profiling in the mouse brain
Cell Reports Methods. 2026-01-01; 6(1): 101267
DOI: 10.1016/j.crmeth.2025.101267
Kapoor S(1), Erni A(1), Vincenzi F(1), Tessier B(2), Venugopal V(2), Meister
G(3), Favereaux A(2), Cremer H(1), Beclin C(4).
Author information:
(1)Aix-Marseille Université, Centre National pour la Recherche Scientifique
(CNRS), Institut de Biologie du Développement de Marseille (IBDM), Marseille,
France.
(2)University of Bordeaux, CNRS Interdisciplinary Institute for Neuroscience,
UMR 5297, Bordeaux, France.
(3)Regensburg, Center for Biochemistry (RCB), University of Regensburg,
Regensburg, Germany.
(4)Aix-Marseille Université, Centre National pour la Recherche Scientifique
(CNRS), Institut de Biologie du Développement de Marseille (IBDM), Marseille,
France. Electronic address: .
AGO-APP through the expression of the T6B peptide permits the isolation of
Ago-bound microRNAs (miRNAs). Here, we present the generation and
characterization of two transgenic mouse lines that enable AGO-APP to be
performed in vivo. First, we generated mice for CRE-dependent T6B expression
throughout the cell. Using this line, we performed AGO affinity purification
(AGO-APP) in olfactory bulb (OB) inhibitory interneurons and cerebral cortex
excitatory neurons. Bioinformatic analysis validated the high reproducibility of
the approach. It also demonstrated that, despite global miRNome conservation
between the two cell types, a set of miRNAs, including the miR-200 family and
the miR-183/96/182 cluster, is massively enriched in OB interneurons, which
aligns with previous observations. In the second mouse line, T6B is fused to the
postsynaptic protein PSD95. Isolation of T6B-PSD95 fractions from OB and
cortical neurons identified specific sets of postsynapse-enriched miRNAs. Gene
ontology analyses confirmed that these miRNAs preferentially target mRNAs
related to synaptic functions.
Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.
DOI: 10.1016/j.crmeth.2025.101267
PMCID: PMC12853173
PMID: 41468888 [Indexed for MEDLINE]
Conflict of interest statement: Declaration of interests The authors declare no
competing interests.