[R35] Spatial Principles of Nuclear RNA Organization
Ente: National Institute of General Medical Sciences
Scadenza: 2031-05-31
Importo max: 462.000 EUR
Paese: US
Descrizione
PROEJCT SUMMARY
Mammalian genomes are pervasively transcribed into both short and long non-coding RNAs (ncRNAs) that
regulate essential processes such as splicing, translation, RNA interference, and dosage compensation, while
also serving as architectural elements that shape nuclear organization. Distinct classes of ncRNAs occupy
characteristic nuclear bodies: MALAT1 in speckles, NEAT1 in paraspeckles, snoRNAs in nucleoli. However, we
lack a comprehensive understanding of how ncRNAs nucleate these compartments and how their spatial
distribution adapts during cellular differentiation or environmental stimuli. Existing ensemble assays such as
CLIP-seq, fractionation, and SPRITE have provided key assignments of ncRNAs to nuclear domains, but these
population-averaged methods cannot resolve single-cell or subnuclear heterogeneity. The goal of this research
direction is to establish a spatial imaging framework that reveals the rules governing ncRNA organization within
the nucleus across developmental and environmental contexts. Building on my prior development of spatially
resolved epigenomic and transcriptomic profiling platforms, this project advances three interconnected themes.
Theme 1 will extend RNA MERFISH to profile small ncRNAs by incorporating enzymatic amplification strategies
such as in situ polyadenylation and T7 amplification, enabling spatial mapping of both short and long ncRNAs.
Theme 2 will define the spatial positioning of ncRNAs within nuclear bodies and track how these associations
are remodeled during neuronal differentiation, integrating RNA MERFISH with multiplexed nuclear body protein
imaging and expansion microscopy. Theme 3 will dissect the structural protein and ncRNA localization in Cajal
and histone locus bodies, and probing necessity and sufficiency through knockdown followed by modular protein
domain rescues. This research direction will advance our fundamental understanding of the spatial biology of
nuclear granules and ncRNA interaction potentially informing further understanding of diseases where the
process goes awry.
Istituzione: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
PI: Cheen Euong Ang
Progetto: 1R35GM166361-01
Settori: National Institute of General Medical Sciences
Vai al bando originale
Registrati gratis su Bandolo per trovare bandi compatibili con la tua azienda.