Biology subjects

Ruepp, M.-D.

Publications and source records attributed to Ruepp, M.-D..

16 recordsLinked to original sources

bioRxiv2026

C9ORF72-derived polyGR polypeptides disrupt passive nucleocytoplasmic transport by tuning protein affinity for the nuclear pore barrier

Nucleocytoplasmic partitioning is an essential determinant of eukaryotic cellular function, governed by the nuclear pore complex, a molecular portal filled by a disordered phenylalanine-glycine (FG)-rich phase that governs selective entry and exit. Disruption of nucleocytoplasmic partitioning is seen across many different diseases, including viral infection, cancer, and neurodegeneration. However, what determines whether a given protein mislocalises when nucleocytoplasmic transport is disrupted remains unknown. This question is central to amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD), where cytosolic mislocalisation of the nuclear RNA-binding protein TDP-43 is a defining pathological feature. The most common genetic cause of these diseases is a G4C2 repeat expansion in the gene C9ORF72, which produces aberrant neurotoxic polypeptides that induce nucleocytoplasmic transport defects. Here, we show how the highly toxic poly(glycine-arginine/GR) polypeptide engages the nuclear pore FG-rich selectivity barrier and retunes passive nucleocytoplasmic transport according to client surface chemistry. Using coarse-grained simulations, in vitro FG-phase reconstitution and human cell lines and neurons, we find that polyGR produces a non-linear, biphasic modulation of nuclear pore passage. Proteins with low affinity for the FG phase are unaffected, whereas proteins with higher affinity due to solvent-exposed hydrophobic residues exhibit enhanced transport up to a critical threshold, beyond which highly hydrophobic proteins experience transport suppression, cytoplasmic accumulation and aggregation. Together, these findings establish how disease-associated polypeptides retune the physicochemical rules governing passive nuclear pore transport, leading to biphasic outcomes determined by protein surface chemistry that alter protein compartmentalisation and aggregation. This provides a biophysical mechanism by which polyGR drives selective protein vulnerability to nuclear pore dysfunction in C9ORF72-associated ALS/FTD.

cell biology↗

bioRxiv2026

Breaking β-sheets in FUS prion-like domain preserves phase separation and function but prevents aggregation and toxicity

The RNA-binding protein Fused in Sarcoma (FUS) undergoes phase separation associated with RNA processing. However, the prion-like low complexity (LC) domain of FUS forms solid-like aggregates in neurodegenerative diseases. Whether the formation of {beta}-sheet structure associated with pathology is also physiologically/functionally relevant is debated. Similarly, if mislocalization alone or concomitant aggregation is responsible for FUS gain-of-function toxicity remains to be probed. Here, we introduce {beta}-sheet breaking proline residues into FUS LC with the goal of preventing cross-{beta}-driven aggregation without disrupting essential functions and phase separation. {beta}-sheet-deficient FUS variants maintain native-like global motions, disorder, and phase separation, but no longer show a liquid-to-solid transition (LST). Biochemical partitioning, cellular localization, and auto- and cross-regulatory functions of FUS all remain essentially unchanged. Conversely, FUS-induced neurodegeneration in several Drosophila models is drastically reduced. These findings suggest a strategy for mitigating disease-related toxicity through backbone structure modulation to prevent prion-like domain protein aggregation. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=198 SRC="FIGDIR/small/706410v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@d15f63org.highwire.dtl.DTLVardef@1cd6221org.highwire.dtl.DTLVardef@e58126org.highwire.dtl.DTLVardef@181ec67_HPS_FORMAT_FIGEXP M_FIG C_FIG SUMMARYThe RNA-binding protein Fused in Sarcoma (FUS) undergoes phase separation as part of its physiological function but can aberrantly aggregate into solid-like assemblies in amyotrophic lateral sclerosis and frontotemporal dementia. To dissect the role of {beta}-sheets in both function and pathological transition, we engineered {beta}-sheet-preventing FUS variants via targeted proline residue insertions in the prion-like disordered region. These variants retained native structure, motions, and phase behavior yet showed dramatically reduced aggregation, both as an isolated prion-like domain and in full-length FUS. Crucially, these variants maintained a panel of FUS cellular functions that depend on FUS condensation but prevented FUS toxicity in fly models of neurodegeneration. Our findings implicate {beta}-sheets as key drivers of FUS condensate maturation and neuronal toxicity, highlighting {beta}-sheet modulation as a therapeutic strategy against FUS-related neurodegeneration. HIGHLIGHTSO_LITargeted proline additions disrupt {beta}-sheet formation in FUS without altering native conformations, dynamics, or phase separation behavior C_LIO_LI{beta}-sheet-deficient FUS variants prevent aggregation and liquid-to-solid transitions while retaining key biological functions C_LIO_LIIn vivo models reveal attenuated toxicity of {beta}-sheet-deficient FUS in Drosophila C_LIO_LI{beta}-sheets are identified as central drivers of condensate maturation and neuronal death, offering a therapeutic entry point for modulating prion-like domain pathology C_LI

biochemistry↗

bioRxiv2026

The Structured RNA-binding Domains and Condensation Capacity of FUS Shape its RNA-binding Landscape and Function.

RNA-binding proteins (RBPs) are key regulators of gene expression and often contain intrinsically disordered regions that drive biomolecular condensation. Yet, how condensation affects RBP specificity and function remains poorly defined. Here, we use strategically designed point mutations to selectively impair canonical RNA-binding and condensation of the amyotrophic lateral sclerosis (ALS)-linked RBP Fused in Sarcoma (FUS). Using automated high-content imaging, we show that both properties shape nuclear ribonucleoprotein condensates and govern distinct aspects of FUS function in the DNA damage response. Transcriptome-wide mapping of FUS-RNA interactions reveals that the canonical RNA-binding domains recognise G-rich and C-rich motifs, whereas condensation selectively enhances binding to G-rich and structured sequences, often acting in concert with canonical RNA-binding to control transcriptional programs and splicing decisions. These findings provide a mechanistic framework for how FUS integrates condensation with sequence-specific RNA recognition to orchestrate nuclear organisation, genome stability and RNA metabolism. Furthermore, our rigorously validated mutants offer a platform for future mechanistic dissection of ALS pathogenesis and the development of targeted therapeutic strategies. HighlightsO_LIFUS condensation and RNA-binding can be functionally uncoupled with targeted point mutations C_LIO_LICondensation drives FUS recruitment and assembly of nuclear RNP condensates C_LIO_LIFUS condensation and RNA-binding play distinct roles in the DNA damage response C_LIO_LIThe structured RNA-binding domains confer specificity to G-rich and C-rich motifs C_LIO_LICondensation enhances FUS binding to G-rich, structured RNA elements C_LIO_LIFUS condensation and canonical RNA-binding synergistically regulate gene expression C_LI

molecular biology↗

bioRxiv2025

RNA secondary structures modulate hnRNPH1-mediated alternative splicing of cold-shock protein RBM3

Alternative splicing is regulated not only by sequence-specific RNA-protein interactions, but also by dynamic RNA secondary structures that remodel under varying cellular conditions. hnRNPH1, a G-rich motif-binding protein, promotes exon skipping across numerous transcripts, yet how RNA structure shapes its regulatory activity remains unclear. Building on our previous work showing that cooler temperature enhances hnRNPH1 binding to RBM3 pre-mRNA and drives poison exon (PE) skipping, we now identify RNA G-quadruplex (rG4) remodelling as the principal cis-acting mechanism underlying this temperature-sensitive regulation. Using small-molecule stabilizers and destabilizers, we show that rG4 folding reduces PE skipping, whereas rG4 destabilization enhances hnRNPH1-mediated PE exclusion. These effects are conserved across neuronal and non-neuronal cells. Integrating proteomics, phosphoproteomics, kinase perturbation assays, and candidates emerging from our prior genome-wide CRISPR screen, we further uncover contributions from temperature-sensitive CDC2-like kinase signalling and the RNA helicase DDX5, a known rG4 resolver and an hnRNPH1/hnRNPF co-factor. Together, our findings establish rG4 remodelling as the dominant driver of hnRNPH1-dependent RBM3 splicing, while implicating helicase activity and kinase signalling as modulatory layers that fine-tune this response. More broadly, this work illustrates how environmentally driven RNA structural transitions interface with enzymatic regulators to achieve temperature-responsive control of alternative splicing.

molecular biology↗

bioRxiv2025

U7 small nuclear RNA splice-switching therapeutics for STMN2 and UNC13A in Amyotrophic Lateral Sclerosis

TDP-43 nuclear depletion in amyotrophic lateral sclerosis (ALS) causes de-repression of cryptic exons (CEs) in multiple transcripts, including UNC13A and STMN2, disrupting synaptic transmission and neurite outgrowth. We developed a therapeutic U7 snRNA (tU7) approach that suppresses TDP-43-dependent mis-splicing, restores target gene expression, rescues neuronal functions in human iPSC-derived neurons, and shows target engagement in vivo, positioning tU7-mediated splicing correction as a promising therapeutic strategy for ALS.

neuroscience↗

bioRxiv2025

A validated panel of commercial antibodies for reliable detection of FET proteins.

The FET protein family comprises the highly conserved RNA-binding proteins FUS, EWS and TAF15 which are implicated in RNA metabolism and neurodegenerative diseases such as ALS and FTLD. Despite their structural similarity, reliable detection of individual FET proteins remains challenging due to antibody cross-reactivity and inconsistent localisation patterns reported in the literature. To address this, we systematically evaluated 23 commercially available antibodies for specificity and performance across western blotting, immunocytochemistry, and immunohistochemistry. Using single and double shRNA knockdowns in HeLa cells, we confirmed target engagement and identified antibodies which were specific as well as those with significant cross-reactivity. Immunofluorescence in rat primary neurons and HeLa cells showed antibody and cell type dependent variations in nuclear and cytoplasmic localisation; from these, we identified a subset that demonstrated high-quality, region-specific staining in postmortem human brain tissue. Our findings highlight substantial variability in antibody performance and underscore the need for rigorous validation to ensure reproducibility in FET protein research. We present a validated panel of antibodies suitable for diverse applications, providing a critical resource for studies of FET protein biology and pathology.

neuroscience↗

bioRxiv2025

CRISPR/Cas9-mediated generation of a de novo C9ORF72 knock-in isogenic iPSC cell bank to model ALS and FTD

BackgroundThe pathogenic G4C2 repeat expansion in the C9ORF72 gene is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Studies focused on delineating the underlying perturbed mechanisms resulting from this genetic mutation are often confounded by the heterogeneity present in current disease models, such as patient-derived iPSC lines, with estimations of up to 50% of the variation in iPSC cell phenotypes resulting from inter-individual differences. Isogenic models, in which the pathogenic mutation is introduced into a defined genetic background, offer a powerful approach to isolating mutation-specific effects and enable high-resolution comparison across distinct ALS/FTD-associated mutations. Such models are essential for uncovering convergent disease mechanisms and improving reproducibility in ALS/FTD research. MethodsA two-step scarless CRISPR/Cas9 genome editing strategy was used to generate isogenic human iPSC lines carrying a de novo knock-in of a disease-length G4C2 repeat expansion in the C9ORF72 locus. The resulting lines underwent thorough quality control and were differentiated into lower motor neurons and assessed for the presence of key ALS/FTD pathologies, including changes to C9ORF72 mRNA and protein expression, RNA foci and dipeptide repeat proteins. ResultsTwo C9ORF72 knock-in iPSC lines were generated with 631 and 600 G4C2 repeats, alongside an isogenic genome editing control line. The C9ORF72 G4C2 repeat expansion knock-in iPSC lines exhibit both loss-of-function and gain-of-function pathological features characteristic of ALS/FTD. Compared to the parental wild-type KOLF2.1J line and isogenic (wild-type) CRISPR control line, these exhibit a significant reduction in C9ORF72 mRNA and protein levels, the presence of RNA foci accumulation, and a marked increase in poly(GA) and poly(GP) dipeptide repeat protein levels in iPSCs and motor neurons. ConclusionsThis is one of the first reports of a successful knock-in of the pathogenic C9ORF72 G4C2 repeat expansion into a human iPSC line, establishing a genetically defined and physiologically relevant model of ALS/FTD. These isogenic lines recapitulate both key loss- and gain-of-function disease pathologies, providing a crucial complement to existing patient-derived iPSC banks. By eliminating confounding genetic background variability, these cell lines will enable more precise interrogation of C9ORF72-linked pathomechanisms and offer a robust platform for comparative studies across the ALS and FTD spectrum, mechanistic investigations, and future therapeutic targeting with enhanced translational relevance.

neuroscience↗

bioRxiv2025

Analysing the effect of full-length and C-terminally truncated progranulin on proliferation, colony formation, and migration in HepG2 and U87 cells

Progranulin, the precursor protein to seven and a half distinct granulin motifs (GRNs), has been implicated in a broad range of diseases. Progranulin depletion is one of the most frequent causes for hereditary Frontotemporal Dementia (FTD). On the other hand, elevated progranulin levels have been associated with increased malignancy of many tumours, manifesting in increased cell proliferation, migration, metastasis formation, and reduced sensitivity to chemotherapeutics. While some functions can be unambiguously attributed to either full-length progranulin or one or multiple of the different GRNs, much about the interplay between progranulin and GRNs remains unknown. Here, we aimed to test the effect of progranulin overexpression on cell-based tumorigenicity assays, assessing proliferation, migration, and colony formation, using the hepatocellular carcinoma cell line HepG2 and the glioblastoma cell line U87. We transduced these cells with lentiviral vectors to overexpress full-length progranulin, two different C-terminally truncated progranulin proteins, lacking either the last two or the last four GRNs, or a triple FLAG-tagged maltose binding protein as a control. We observed increased colony formation in HepG2 overexpressing the full-length progranulin but not the C-terminally truncated constructs. The U87 cell lines were neither affected by an increase in progranulin levels nor by the depletion of progranulin.

cell biology↗

bioRxiv2025

Generation of C9orf72 repeat knock-in iPSC lines for modelling ALS and FTD

Induced pluripotent stem cell (iPSC) models are powerful tools for neurodegenerative disease modelling, as they allow mechanistic studies in a human genetic environment and they can be differentiated into a range of neuronal and non-neuronal cells. However, these models come with inherent challenges due to line-to-line and clonal variability. To combat this issue, the iPSC Neurodegenerative Disease Initiative (iNDI) has generated an iPSC repository using a single clonal reference line, KOLF2.1J, into which disease-causing mutations and revertants are introduced via gene editing. Here we describe the generation and validation of lines carrying the most common causative mutation for amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), a repeat expansion in the C9orf72 gene, for the iNDI collection of neurodegenerative iPSC models. We demonstrate that these C9orf72 knock-in lines differentiate efficiently into neurons and display characteristic C9orf72-associated pathologies, including reduced C9orf72 levels and the presence of dipeptide repeat proteins (DPRs) and RNA foci, which increase in abundance over time in culture. These pathologies are not present in revertant cells lacking the repeat expansion. These repeat expansion and revertant cell lines are now available to academic and for-profit institutions through the JAX iPS cell repository and will help to facilitate and standardise iPSC-based ALS/FTD research.

neuroscience↗

bioRxiv2023

Single cell RNA sequencing in isogenic FUS and TARDBP mutant ALS lines reveals early mitochondrial dysfunction as a common pathway in motor neurons

Mutations in the RNA/DNA-binding proteins FUS and TDP-43 cause the fatal disease amyotrophic lateral sclerosis (ALS). The precise mechanisms behind the selective motor neuron degeneration remain unclear and it is uncertain if ALS-causative mutations trigger motor neuron death through shared or distinct pathogenic pathways. To address these two questions, we performed single-cell RNA sequencing across neuron types derived from isogenic induced pluripotent stem cell lines, harbouring FUS P525L, FUS R495X, TARDBP M337V mutations or FUS knockout. The mutations elicited 5- to 15-fold greater transcriptional responses in motor neurons than interneurons. Approximately 20% of transcripts uniquely dysregulated in motor neurons were shared across FUS mutations, with half being driven by FUS gain-of-function. Among these, a majority pointed towards mitochondrial impairments, with attenuated pathways shared with the TARDBP M337V mutation. Meta-analysis demonstrated convergence on mitochondrial dysfunction with C9orf72-ALS patient-derived motor neurons. We observed impaired mitochondrial motility across ALS motor axons, even in isogenic FUS R244C motor neurons, which retain FUS in the nucleus, demonstrating shared toxic gain-of-function mechanisms across FUS- and TARDBP-ALS, uncoupled from protein mislocalization. These early signs of mitochondrial dysfunction unique to motor neurons could have profound implications for their survival and represent promising therapeutic targets across multiple ALS forms.

neuroscience↗

bioRxiv2022

HNRNPH1 regulates the neuroprotective cold-shock protein RBM3 expression through poison exon exclusion

Enhanced expression of the cold-shock protein RNA binding motif 3 (RBM3) is highly neuroprotective both in vitro and in vivo. Whilst upstream signalling pathways leading to RBM3 expression have been described, the precise molecular mechanism of RBM3 induction during cooling remains elusive. To identify temperature-dependent modulators of RBM3, we performed a genome-wide CRISPR-Cas9 knockout screen using RBM3-reporter human iPSC-derived neurons. We found that RBM3 mRNA and protein levels are robustly regulated by several splicing factors, with heterogeneous nuclear ribonucleoprotein H1 (HNRNPH1) being the strongest positive regulator. Splicing analysis revealed that moderate hypothermia significantly represses the inclusion of a poison exon, which, when retained, targets the mRNA for nonsense-mediated decay. Importantly, we show that HNRNPH1 mediates this cold-dependent exon skipping via its interaction with a G-rich motif within the poison exon. Our study provides novel mechanistic insights into the regulation of RBM3 and provides further targets for neuroprotective therapeutic strategies. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/514062v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@ef0a5org.highwire.dtl.DTLVardef@d907a5org.highwire.dtl.DTLVardef@85176aorg.highwire.dtl.DTLVardef@1c4306a_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2022

The cancer-associated RBM39 bridges the pre-mRNA, U1 and U2 snRNPs to regulate alternative splicing

Pharmacologic depletion of RNA-binding motif 39 (RBM39) using aryl sulfonamides represents a promising anti-cancer therapy. However, its efficiency correlates with the expression level of DCAF15 which acts at the interface between RBM39, the drug and the E3-ubiquitin ligase. Consequently, the identification of alternative approaches to deplete RBM39 independently of DCAF15 is required. Here, we combined transcriptomic analysis, functional assays, and structural biology to elucidate the molecular mechanisms governing RBM39 homeostasis. Our data revealed that RBM39 autoregulates the splicing of its own pre-mRNA by triggering the inclusion of a poison exon. During this process, RBM39 selects the 3-splice site of the toxic exon, helps the recruitment of U1 snRNP on its weak 5-splice site and bridges the 3-splice site recognition machinery. The elucidation of the molecular mechanisms controlling RBM39 homeostasis provides unprecedented insights into alternative 3-splice site selection and a solid frame to design alternative anti-cancer therapies.

molecular biology↗

bioRxiv2019

The phase separation-dependent FUS interactome reveals nuclear and cytoplasmic function of liquid-liquid phase separation

Liquid-liquid phase separation (LLPS) of proteins and RNAs has emerged as the driving force underlying the formation of membrane-less organelles. Such biomolecular condensates have various biological functions and have been linked to disease. One of the best studied proteins undergoing LLPS is Fused in Sarcoma (FUS), a predominantly nuclear RNA-binding protein. Mutations in FUS have been causally linked to Amyotrophic Lateral Sclerosis (ALS), an adult-onset motor neuron disease, and LLPS followed by aggregation of cytoplasmic FUS has been proposed to be a crucial disease mechanism. In spite of this, it is currently unclear how LLPS impacts the behaviour of FUS in cells, e.g. its interactome. In order to study the consequences of LLPS on FUS and its interaction partners, we developed a method that allows for the purification of phase separated FUS-containing droplets from cell lysates. We observe substantial alterations in the interactome of FUS, depending on its biophysical state. While non-phase separated FUS interacts mainly with its well-known interaction partners involved in pre-mRNA processing, phase-separated FUS predominantly binds to proteins involved in chromatin remodelling and DNA damage repair. Interestingly, factors with function in mitochondria are strongly enriched with phase-separated FUS, providing a potential explanation for early changes in mitochondrial gene expression observed in mouse models of ALS-FUS. In summary, we present a methodology that allows to investigate the interactome of phase-separating proteins and provide evidence that LLPS strongly shapes the FUS interactome with important implications for function and disease.

molecular biology↗

bioRxiv2019

FUS-dependent phase separation initiates double-strand break repair

RNA-binding proteins (RBPs) are emerging as important effectors of the cellular DNA damage response (DDR). The RBP FUS is implicated in RNA metabolism and DNA repair, and it undergoes reversible liquid-liquid phase separation (LLPS) in vitro. Here, we demonstrate that FUS-dependent LLPS is necessary for the initiation of the DDR. Using laser microirradiation in FUS-knockout cells, we show that FUS is required for the recruitment to DNA damage sites of the DDR factors KU80, NBS1, 53BP1, and of SFPQ, another RBP implicated in the DDR. The relocation of KU80, NBS1, and SFPQ is similarly impaired by LLPS inhibitors, or LLPS-deficient FUS variants. We also show that LLPS is necessary for efficient {gamma}H2AX foci formation. Finally, using super-resolution structured illumination microscopy, we demonstrate that the absence of FUS impairs the proper arrangement of {gamma}H2AX nano-foci into higher-order clusters. These findings demonstrate the early requirement for FUS-dependent LLPS in the activation of the DDR and the proper assembly of DSBs repair complexes.

molecular biology↗

bioRxiv2019

FUS ALS-causative mutations impact FUS autoregulation and the processing of RNA-binding proteins through intron retention

Mutations in the RNA-binding protein FUS cause amyotrophic lateral sclerosis (ALS), a devastating neurodegenerative disease in which the loss of motor neurons induces progressive weakness and death from respiratory failure, typically only 3-5 years after onset. FUS plays a role in numerous aspects of RNA metabolism, including mRNA splicing. However, the impact of ALS-causative mutations on splicing has not been fully characterised, as most disease models have been based on FUS overexpression, which in itself alters its RNA processing functions. To overcome this, we and others have recently created knock-in models, and have generated high depth RNA-sequencing data on FUS mutants in parallel to FUS knockout. We combined three independent datasets with a joint modelling approach, allowing us to compare the mutation-induced changes to genuine loss of function. We find that FUS ALS-mutations induce a widespread loss of function on expression and splicing, with a preferential effect on RNA binding proteins. Mutant FUS induces intron retention changes through RNA binding, and we identify an intron retention event in FUS itself that is associated with its autoregulation. Altered FUS regulation has been linked to disease, and intriguingly, we find FUS autoregulation to be altered not only by FUS mutations, but also in other genetic forms of ALS, including those caused by TDP-43, VCP and SOD1 mutations, supporting the concept that multiple ALS genes interact in a regulatory network.

neuroscience↗

bioRxiv2018

FUS controls the processing of snoRNAs into smaller RNA fragments that can regulate gene expression

FUS is a multifunctional protein involved in many steps of RNA metabolism, including transcription, splicing, miRNA processing and replication-dependent histone gene expression. In this paper, we show for the first time that FUS binds and negatively regulates the levels of a subset of snoRNAs in cells. Scanning of available human small RNA databases revealed the existence of smaller RNA fragments that can be processed from FUS-dependent snoRNAs. Therefore, we suggest that FUS mediates the biogenesis of snoRNA-derived small RNAs, called sdRNAs. Further in silico approaches enabled us to predict putative targets of selected FUS-dependent sdRNAs. Our results indicate that sdRNAs may bind to different regions of target mRNAs as well as to noncoding transcripts and influence the posttranscriptional level or translation of these targets. SIGNIFICANCE STATEMENTRNA metabolism is orchestrated by a complex network of RNA-protein interactions and involves various classes of RNA molecules. Small nucleolar RNAs (snoRNAs) are commonly considered essential components of the ribosome biogenesis pathway. However, recent studies have revealed that snoRNAs can also be fragmented into small entities called snoRNA-derived RNAs (sdRNAs), which have been linked to multiple cancer types and thus may serve as next-generation prognostic or diagnostic biomarkers. In this paper, a multifunctional protein, FUS, was shown to be involved in the biogenesis of snoRNA-derived fragments. Furthermore, we combined bioinformatic analyses with complementary experimental approaches to elucidate the role of FUS-dependent sdRNAs in gene expression regulation. Our findings reveal the considerable regulatory potential of this new class of small noncoding RNAs.

molecular biology↗