Myotonic dystrophy type 1 is the most prevalent adult-onset muscular dystrophy and is characterized by progressive muscle weakness, myotonia, cardiac conduction defects, endocrine dysfunction, and central nervous system involvement. Myotonic dystrophy type 1is caused by an unstable CTG repeat expansion in the 3' untranslated region of the DMPK gene, which produces toxic CUG-expanded transcripts that sequester RNA-binding proteins such as Muscleblind-like, induce widespread alternative splicing defects, and drive an RNA gain-of-function mechanism rather than simple DMPK haploinsufficiency. Despite major advances in understanding the molecular pathogenesis of myotonic dystrophy type 1, there is still no approved cure or disease-modifying therapy. This review summarizes the molecular basis of myotonic dystrophy type 1 and provides an in-depth overview of emerging therapeutic strategies that directly target the underlying pathogenic cascade at the DNA and RNA levels. Gene therapy-based approaches, including CRISPR-mediated genome editing, aim to reduce or eliminate the expanded CTG repeats or expanded DMPK allele and its toxic transcripts. In parallel, a broad spectrum of RNA-directed interventions is being developed, encompassing antisense oligonucleotides, antibody-penetrating and cell-penetrating peptide-conjugated antisense oligonucleotides to enhance skeletal and cardiac muscle delivery, small interfering RNAs, and microRNA-based tools such as antagomiRs. Additional strategies exploit engineered RNA-binding proteins and peptide decoys to disrupt toxic ribonuclear aggregates, polyadenylation signal-driven premature transcriptional termination to selectively silence mutant DMPK, and small molecules that modulate RNA metabolism, dissolve CUG RNA foci, or correct downstream mis-splicing. By integrating data from preclinical models and ongoing clinical trials, including recent advances with muscle‑targeted antisense oligonucleotide conjugates and gene therapy, this review outlines the current status, strengths, and limitations of these mechanism-based therapies for myotonic dystrophy type 1. The discussion highlights key translational challenges such as efficient delivery to skeletal muscle, the heart, and brain, long-term safety, and robust pharmacodynamic biomarkers as well as opportunities for combination and next-generation approaches aimed at converting molecular correction into durable clinical benefit for patients with myotonic dystrophy type 1.

Escherichia coli is an ideal microbial chassis for producing high-value compounds due to its well-characterized genome and extensive genetic toolbox. Shikimic acid, an intermediate in the aromatic amino acid pathway and the main precursor for oseltamivir, a drug used to treat influenza, is of considerable industrial importance. Here, we present a proof-of-concept metabolic control strategy that combines a biphasic control module with antisense RNA (asRNA) to redirect carbon flux toward shikimic acid accumulation. An asRNA targeting the aroL gene was designed to include the MicF M7.4 Hfq-binding motif and a complementary region targeting the translation initiation sequence of aroL. The construct was expressed from the PBAD promoter in E. coli JW5947-1 (ΔaroK) using the plasmid pBR322. The biphasic system enables staged control of carbon partitioning. In the absence of the PBAD inducer, antisense RNA expression is repressed, allowing shikimic acid utilization for aromatic amino acid synthesis. Upon induction, antisense RNA expression represses shikimate kinase II and promotes shikimic acid accumulation. Shake-flask fermentations showed that extracellular shikimic acid increased to 12.86 ± 1.70 mg L-1 compared to 6.20 ± 0.68 mg L-1 in the control. Yield increased (∼2.6-fold) to 157.1 μg/g (±19 μg/g) after induction compared to 82.9 μg/g (±17 μg/g) prior to induction. This work introduces a portable, nongenome-editing, trans-acting regulatory module that enables biphasic flux control and can be integrated with chromosomal or transporter engineering for scalable production.

Gene delivery for cerebral neurodegenerative disorders - current advancements and limitations.

Prostate cancer (PCa) ranks as the fifth most prevalent cancer type globally, with its incidence rising due to lifestyle changes. The prognosis significantly worsens at the metastatic stage, with a 5-year survival rate dropping to 30%. This study focuses on the androgen signaling pathway's role in PCa etiology and aims to identify molecular mechanisms that could serve as potential biomarkers for early diagnosis of the disease in a population of Iranian men. We analyzed the expression levels of long non-coding RNAs (lncRNAs) PCAT18, CTBP1 antisense RNA (CTBP1-AS), and HOTAIR in blood samples from 30 PCa patients and 30 individuals with benign prostatic hyperplasia (BPH). RT-PCR and specific primers were used for quantification. We observed a significant downregulation of PCAT18, HOTAIR and CTBP1-AS in the PCa cohort compared to BPH. ROC curve analysis revealed HOTAIR as the most promising biomarker, with an area under the curve of 0.6897. Further analysis indicated that PCAT18 and HOTAIR demonstrate considerable sensitivity and specificity for distinguishing PCa from BPH, suggesting their potential utility in clinical diagnosis and prognosis evaluation. Significant correlation was found between LNCRNAs expression and demographic information for CTBP1-AS expression vs. size of tumor, PCAT18 expression vs. PSA and PCAT18 expression vs. grade. These results implying that lncRNAs dysregulation may be more indicative of cancer biology. These findings highlight the importance of CTBP1, PCAT18 and HOTAIR as potential biomarkers in prostate cancer and underscore the necessity for further investigation into their roles in cancer progression and therapeutic strategies.

Transcriptional interference is rarely documented in bacteria. In cyanobacteria, phycobilisomes are the major light-harvesting antennae, and their degradation under non-optimal conditions follows a tightly regulated genetic program leading to the production of NblA, a widely conserved proteolytic adapter. NblA production is regulated at both the transcriptional and post-transcriptional levels. Here, we uncover an additional regulatory layer mediated by an antisense RNA conserved in heterocyst-forming cyanobacteria. In Nostoc sp. PCC 7120, transcription of the abundant antisense RNA (as_nblA) limits nblA mRNA accumulation. A strain unable to transcribe as_nblA produces an excess of NblA, ultimately so harmful that suppressor mutations of nblA expression accumulate rapidly, underscoring the essential role of as_nblA. Rifampicin time-series experiments and the inability of as_nblA to regulate nblA expression in trans support transcriptional interference as the primary regulatory mechanism of as_nblA. Mathematical modeling of nblA expression, supported by biological data, shows that as_nblA plays a pivotal role in preventing leaky nblA expression under non-inducing conditions. Our work highlights the importance of antisense RNA-mediated regulation, particularly transcriptional interference, in establishing thresholds that prevent spurious expression of genes encoding critical cellular functions.

Riboregulation is a key component of bacterial adaptation, as regulatory RNAs enable rapid and precise control of gene expression in response to stress and fluctuating environmental conditions. This control is exerted at the levels of transcription, translation, and mRNA stability, allowing the cell to efficiently adjust gene expression without the need to produce additional protein regulators. Regulation involves both locally acting elements (including riboswitches, thermosensors, and antisense RNAs) and in trans acting molecules primarily sRNAs which form extensive networks affecting the expression of many genes simultaneously. The multilayered nature of these systems is further enhanced by sRNA-binding proteins and RNA sponges, which modulate the availability of regulators and shape complex gene expression control networks. Modern methods like Hfq-CLASH enable better identification of these interactions in vivo. In parallel, applied research leverages these mechanisms in synthetic biology and against antibiotic resistance.

Inflammasome-related markers and long non-coding rnas in seminal plasma: Associations with sperm DNA fragmentation and male infertility.

Updates on hereditary transthyretin amyloidosis polyneuropathy.

The Arabidopsis enolase2 (AtENO2) gene produces both the glycolytic enzyme enolase and an N-terminal truncated isoform, termed the transcriptional repressor Arabidopsis cMyc-Binding Protein 1 (AtMBP-1), through alternative translation. Maintaining AtENO2 homeostasis via feedback repression by AtMBP-1 is essential for its biological functions; however, the underlying regulatory mechanism remains unclear. Cadmium (Cd), a toxic soil pollutant, strongly inhibits plant growth and development. Interestingly, Cd induces enolase accumulation in plants, and AtMBP-1 binds to metal-responsive elements (MREs) that confer a Cd response in Arabidopsis. Here, we demonstrate that AtMBP-1-mediated repression via an MRE in the AtENO2 promoter maintains the AtENO2 expression balance, which is required for Arabidopsis Cd resistance. Knockdown of AtENO2 via antisense RNA increased Cd sensitivity, which was accompanied by altered intermediates of the tryptophan pathway (notably those involved in auxin biosynthesis), elevated Cd accumulation in the shoots and roots, and increased lipid peroxidation. We further showed that AtENO2 utilizes its 5'-UTR as a Cd-responsive alternative promoter, driving a predominant short transcript that is efficiently translated into the AtENO2 protein. Accordingly, Cd stress increased AtENO2 protein accumulation. An MRE located within the 5'-UTR promoter was functional: CRISPR/Cas9-mediated deletion of a 50 bp MRE-containing fragment in vivo increased AtENO2 abundance and significantly enhanced Arabidopsis growth under Cd stress. AtMBP-1 binds to this MRE to repress AtENO2 promoter activity, thereby forming an autoregulatory loop that fine-tunes AtENO2 levels and prevents its overaccumulation during prolonged Cd stress. Our findings reveal a previously unknown mechanism underlying AtENO2 function in plant adaptation to heavy metal stress.

Lung cancer is a leading cause of cancer mortality, with non-small cell lung cancer (NSCLC) comprising the majority of cases. This study aims to investigate the functional role of long non-coding RNA (lncRNA) PTPRG antisense RNA 1 (PTPRG-AS1) in NSCLC progression using in vitro models, focusing on its potential as a regulator of key oncogenic processes and its interaction with the IGF-1/PI3K/AKT signaling pathway to identify novel therapeutic targets. PTPRG-AS1 expression in NSCLC cell lines (A549, H1299, NCI-H226) and normal lung cells (Beas-2B) was analyzed using RT-qPCR. PTPRG-AS1 was silenced with siRNA, and its effects on cell proliferation, migration, invasion, colony formation, apoptosis, and angiogenesis-related factors were evaluated. Western blot analysis assessed components of the PI3K/AKT pathway. LncRNA PTPRG-AS1 was significantly upregulated in NSCLC cells, particularly in H1299. Our results showed that PTPRG-AS1 knockdown inhibited cell proliferation, migration, and invasion, while promoting cell apoptosis. IGF-1 treatment reversed these effects. PTPRG-AS1 knockdown reduced levels of angiogenesis-related factors, including VEGF and bFGF. Additionally, silencing PTPRG-AS1 decreased expression of key PI3K/AKT pathway components, while exogenous IGF-1 reactivated this signaling cascade. PTPRG-AS1 plays a critical role in NSCLC progression by modulating the IGF-1/PI3K/AKT signaling pathway. Targeting PTPRG-AS1 offers a promising therapeutic strategy for NSCLC, warranting further investigation into its clinical implications.

Deciphering the directed iron-stress regulatory network of Synechocystis sp. PCC 6803 reveals novel master regulators.

BackgroundCervical cancer (CESC) remains a major global health burden, and its molecular mechanisms are not fully understood. Long non-coding RNAs (lncRNAs) have emerged as critical regulators in tumor biology. Among them, Wilms tumor 1 antisense RNA (WT1-AS) has been implicated in several cancers, but its role in CESC is largely unknown. Therefore, this study aimed to investigate the expression pattern, prognostic significance, biological function, and upstream transcriptional regulation of WT1-AS in CESC.MethodsWe performed a comprehensive analysis integrating The Cancer Genome Atlas (TCGA) pan-cancer and CESC datasets to examine the expression profile and prognostic relevance of WT1-AS. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and single-cell RNA sequencing analyses were conducted to explore its biological functions. Functional assays, including Western blotting, flow cytometry, and xenograft mouse models, were performed to validate the effects of WT1-AS on apoptosis and tumor growth. Upstream transcriptional regulation was investigated using transcription factor prediction, co-expression analysis, and luciferase reporter assays.ResultsWT1-AS was significantly dysregulated across multiple cancers and correlated with poor prognosis in several tumor types. In CESC, WT1-AS was primarily expressed in malignant epithelial cells and fibroblasts and was functionally enriched in apoptosis- and extracellular matrix-related pathways. Overexpression of WT1-AS promoted apoptosis, as evidenced by increased cleaved-caspase3 and cleaved-PARP expression, and suppressed tumor growth in vivo. Mechanistically, odd-skipped related transcription factor 2 (OSR2) was identified as a direct transcriptional regulator of WT1-AS. Overexpression of OSR2 enhanced apoptosis, and co-overexpression with WT1-AS further amplified pro-apoptotic effects.ConclusionsOur findings reveal that WT1-AS functions as a tumor suppressor in CESC by promoting apoptosis and identify OSR2 as a novel upstream regulator. The WT1-AS/OSR2 axis may have biological and potential prognostic relevance in CESC, although its clinical applicability requires further validation.

The myotonic dystrophy type 1 drug development pipeline: 2026 Edition.

Phelan-McDermid syndrome (PMS) is caused in the majority of cases by the loss or mutation of one allele of the SHANK3 gene, located in human chr 22q13.33. PMS displays large interindividual differences in clinical severity and longitudinal trajectory. Other genes located in this chromosomal region are known to contribute to the clinical phenotype (CELSR1, TCF20) in patients with larger deletions. The aim of this study is to identify clinically-relevant phenotypic features significantly influenced by the size of chromosome 22q terminal deletion and to identify new potential candidate genes likely to be involved in these phenotypic effects. Genotype-phenotype correlations were investigated in 63 PMS patients directly ascertained by deep clinical phenotyping and determination of deletion size (Agilent CGH-array 180K or 400K). Patients were partitioned into eleven categories, based on deletion size (Mb). Phenotypic variables significantly influenced by deletion size were initially detected by exact χ2 (10,000 iterations) and Kendall's Tau. Candidate genes were then sought using: (a) ROC curves for binary dichotomous variables; (b) best separation threshold for quantitative variables. Phenotypic variables significantly associated with chromosome 22q deletion size in our sample include: expressive language (p < 0.001); motor development timing (p < 0.001); gait (p < 0.001); muscle strength (p < 0.01); social cognition, encompassing eye contact, exchange gesture, and joint attention (p < 0.001-< 0.05); infectious diseases coincident with the onset of behavioral manifestations (p < 0.001); brain structural abnormalities on MRI (p < 0.001); dysmorphisms (p < 0.001); renal and urinary malformations (p < 0.01); comorbid lifelong bipolar disorder (p < 0.05). The best separation thresholds for many of these variables were located within or nearby genes playing important morphogenetic (PLXNB2, TAFA5) or neurodevelopmental roles (BRD1, TBC1D22A, ATXN10 and/or FBLN1). For renal malformations, the two best thresholds point toward one long non-coding RNA and a cluster of antisense RNAs. The genes identified in this study appear as strong candidates to contribute to the PMS phenotype, by conferring an additional layer of abnormal neurodevelopment and impaired morphogenesis to the disruptive effects produced by SHANK3 haploinsufficiency.

Potential use of HOTAIR and MALAT1 as prognostic biomarkers for cervical cancer: An application to the NHS.

Genomes constantly face threats from transposable elements (TEs) and other genomic parasites. While the silencing of existing TE has been well studied, little is known about how cells recognize new invading TEs that they have not previously encountered. Here we explore this question by inserting foreign sequences into S. pombe. Our data revealed that the newly invading TE tj1 is recognized and targeted for silencing by RNAi and heterochromatin. The efficiency of recognition, as well as the degree and stability of silencing, depends on the copy number and insertion location of the TE. We demonstrated that RNA, rather than DNA, is sensed, and that the efficiency of TE recognition correlates with levels of RNA antisense to the TE, generated from upstream transcripts. We also show that various genes of non-transposable nature can initiate silencing. Our data show that silencing may not require recognition of specific elements in the transposon by the host defense systems, and suggest that disruption of host transcription patterns triggers recognition of TE.

Pseudoalteromonas haloplanktis TAC125 is a psychrophilic marine bacterium widely used to study cold adaptation and increasingly exploited as a non-conventional platform for biotechnological applications. The strain harbors the endogenous megaplasmid pMEGA (64.7 kb), whose presence may limit its exploitation as a cell factory, making its elimination advantageous to strain engineering. Traditional plasmid-curing approaches based on chemical and physical agents are often inefficient and unsuitable for stable endogenous replicons, such as pMEGA. Here, we describe a targeted protocol for pMEGA curing in P. haloplanktis TAC125 that combines homologous recombination with paired-termini antisense RNA (PTasRNA) gene silencing. First, a selectable marker cassette is inserted into pMEGA by homologous recombination using a suicide vector, enabling selective discrimination between plasmid-positive and plasmid-cured bacteria. Next, PTasRNA gene silencing technology is applied to target a gene essential for the replication of pMEGA, thereby transiently interfering with its replication and promoting its loss. This approach provides a specific method to cure a highly stable endogenous megaplasmid in a psychrophilic non-conventional bacterium, enabling improved functional studies and strain optimization, establishing a broadly applicable framework for targeted curing across diverse bacterial systems.Key features• Enables targeted curing of stable endogenous plasmids lacking selectable markers.• Combines replication silencing and homologous recombination for targeted plasmid elimination without permanent chromosomal modification.• Adaptable framework for non-model bacteria with limited genetic toolkits, such as marine psychrophiles.

Oral squamous cell carcinoma (OSCC) remains a major global health challenge due to late diagnosis, high recurrence, and limited therapeutic success. This review focuses on circulating long non-coding RNAs (lncRNAs) as emerging biomarkers and therapeutic targets that can transform the clinical management of OSCC. Unlike earlier studies restricted to tissue-based analyses, this work emphasizes circulating lncRNAs detectable in saliva, plasma, and serum, highlighting their non-invasive diagnostic potential and mechanistic relevance in tumor progression, metastasis, and therapy resistance. Relevant studies published up to 2025 were systematically reviewed from PubMed and Google Scholar to synthesize current knowledge on their biogenesis, secretion through extracellular vesicles and RNA-binding proteins, detection technologies, and clinical applicability. Several circulating lncRNAs, including HOTAIR, MALAT1, MEG3, ANRIL, and LINC00657, show strong associations with tumor stage, prognosis, and therapeutic outcomes. The review also discusses emerging RNA-based therapeutic approaches such as antisense oligonucleotides, RNA interference, and CRISPR-based gene editing. Overall, the analysis underscores the diagnostic precision, prognostic power, and therapeutic promise of circulating lncRNAs, proposing them as key components in the future of liquid biopsy-driven, personalized oncology for OSCC.

Diabetes mellitus (DM) with coronary artery disease (CAD), referred to as DM-CAD, is a prevalent endocrine-metabolic condition characterized by macrophage-driven inflammation and metabolic dysregulation. This study aims to investigate the role of YTH N6-methyladenosine RNA binding protein F1 (YTHDF1)-mediated N6-methyladenosine (m6A) modification in stabilizing the long non-coding RNA (lncRNA) Opa interacting protein 5 antisense RNA 1 (OIP5-AS1) and to determine its impact on macrophage metabolic dysfunction in DM-CAD. Single-cell RNA sequencing and bulk RNA sequencing were performed using DM-CAD mouse models, with downstream analyses conducted using Seurat, CellChat, least absolute shrinkage and selection operator (LASSO) regression, and random forest algorithms. Experimental validation included RNA immunoprecipitation followed by quantitative polymerase chain reaction, actinomycin D-based RNA stability assays, and 2-deoxyglucose (2-DG) interventions in THP-1-derived macrophages, alongside metabolomic profiling and reactive oxygen species (ROS) measurements. Increased macrophage-endothelial cell coupling was observed in DM-CAD, with both OIP5-AS1 and YTHDF1 significantly upregulated and closely associated with glycolytic metabolic pathways. YTHDF1-mediated m6A modification stabilized OIP5-AS1, thereby promoting glycolysis, foam cell formation, ROS production, plaque development, and the upregulation of proinflammatory cytokines, collectively exacerbating atherosclerosis. Notably, treatment with 2-DG markedly reversed these pathological phenotypes. This study identifies the YTHDF1-m6A-OIP5-AS1 axis as a critical regulator of macrophage metabolic dysfunction in DM-CAD, thereby providing an epigenetic framework for understanding disease progression. Targeting this regulatory pathway may attenuate metabolic inflammation and represents a promising therapeutic strategy for endocrine-related cardiovascular complications.

Long non-coding RNAs (lncRNAs) play key roles in gene regulation. One potential regulation mechanism involves the formation of RNA•DNA:DNA triplexes. In these triplexes, the lncRNA binds in the major groove of a target DNA via Hoogsteen base pair formation. Here, we investigated the impact of the underlying RNA binding on the on the stability of the DNA duplex target to gain insights into the triplex stability at base pair resolution. Quantification of the temperature-dependent exchange of imino hydrogen atoms with solvent of the target DNA duplex allows determination of the changes of the stability of individual DNA duplex base pairs upon triplex formation. The data shown here investigates an antiparallel triplex, formed between the lncRNA hypoxia-inducible factor 1-alpha Antisense RNA 1 (HIF1α-AS1) and the DNA target Adrenomedullin (ADM), important in cardiovascular diseases. Triplex formation alters DNA structure and stability by affecting both hydrogen bonding strength and nucleobase-stacking interactions. These thermodynamic insights support bioinformatic methods to predict triplex stability and enhance our understanding of RNA•DNA:DNA triplex formation.