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Mapping microRNA-target interactions in abundant cell types with Spy3-AGO2 Pull-down [SAP-seq]

ABSTRACT: To illuminate the molecular mechanisms driving neuronal differentiation we generated a mouse line amenable to mapping miRNA-target interactions in rare cell types. Biochemical approaches to purify AGO2-miRNA-target complexes have successfully mapped MTIs in abundant populations of neurons. However, due to their technical complexity and high background, these approaches are not suitable for mapping interactions in rare cell populations such the many neuronal subtypes that compose the mammalian brain. We therefore generated a mouse line with a conditional SpyTag3, which is small and offers near-infinite affinity for pull-downs, in the endogenous Ago2 gene. We then developed a method Spy3-AGO2 pull-down and sequencing (SAPseq), which we first benchmarked for in vivo use using cortical pyramidal neurons, an abundant population.

ORGANISM(S): Mus musculus

PROVIDER: GSE289640 | GEO | 2025/04/02

REPOSITORIES: GEO

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Mapping microRNA-target interactions in rare cell types with Spy3-AGO2 Pull-down

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