Dottore di ricerca

ciclo: XXXIII

Titolo della tesi: The long non-coding RNA lncMN2 functions as a competing endogenous RNA in motor neuron differentiation

Long non-coding RNAs (lncRNAs) are currently recognized as crucial players in nervous system development, function and pathology. In this study we used an in-vitro differentiation system to generate mouse motor neurons (MNs) from embryonic stem cells and we analyzed, via RNA-Seq, the long non-coding transcriptome of MNs. We selected 12 species of lncRNAs based on their expression levels and fold change along MN differentiation. Among these candidates, I focused my attention on lncRNA 5330434G04Rik (named lncMN2) highlighting peculiar structural and localization features. Functional characterization of the selected lncRNA was performed through CRISPR/Cas9 – KO approach. Cytofluorimetric and phenotypical analysis reveal an impaired motor neuronal differentiation in the lncMN2 – KO clones. Moreover, bioinformatics predictions highlighted a 100nt region on the lncRNA able to bind miR-466i, afterwards confirmed by CLIP and reporter assays, imputing a role to lncMN2 as a ceRNA. Functional characterization of the miRNA binding region was performed through CRISPR/Cas9 – KO approach. Cytofluorimetric and phenotypical analysis on mir-466i binding region – KO clones resembled the already-observed lncMN2-KO phenotypes. Putative miRNA targeted transcripts, underlying the phenotypical evidences, were highlighted through scRNA-seq and confirmed through molecular analysis. Our work aims to investigate lncRNA functions in motor neuronal commitment pathways and is a step toward the comprehension of the still poorly-characterized ceRNA mechanisms in motoneuronal circuitries.

Produzione scientifica

11573/1649233 - 2022 - A multifunctional locus controls motor neuron differentiation through short and long noncoding RNAs
Carvelli, A.; Setti, A.; Desideri, F.; Galfre, S. G.; Biscarini, S.; Santini, T.; Colantoni, A.; Peruzzi, G.; Marzi, M. J.; Capauto, D.; Di Angelantonio, S.; Ballarino, M.; Nicassio, F.; Laneve, P.; Bozzoni, I. - 01a Articolo in rivista
rivista: EMBO JOURNAL (attuale: NATURE PUBLISHING GROUP, 345 PARK AVENUE SOUTH, NEW YORK, USA, NY, 10010-1707 Precedente: Oxford University Press:Journals Department, Great Clarendon Street, Oxford OX2 6DP United Kingdom:011 44 1865 556767, EMAIL:, INTERNET:, Fax: 011 44 1865 267485) pp. - - issn: 0261-4189 - wos: WOS:000810290700001 (6) - scopus: 2-s2.0-85131718309 (6)

11573/1613888 - 2021 - West Nile and Usutu viruses co-circulation in central Italy: outcomes of the 2018 integrated surveillance
Scaramozzino, P.; Carvelli, A.; Bruni, G.; Cappiello, G.; Censi, F.; Magliano, A.; Manna, G.; Ricci, I.; Rombola, P.; Romiti, F.; Rosone, F.; Sala, M. G.; Scicluna, M. T.; Vaglio, S.; De Liberato, C. - 01a Articolo in rivista
rivista: PARASITES & VECTORS (London: BioMed Central-Springer Nature.) pp. 243-249 - issn: 1756-3305 - wos: WOS:000656498300004 (13) - scopus: 2-s2.0-85105459430 (13)

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