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Alternative polyadenylation (APA) creates distinct transcripts from the same gene by cleaving the pre-mRNA at poly(A) sites that can lie within the 3′ untranslated region (3′UTR), introns, or exons. Most studies focus on APA within the 3′UTR; however, here, we show that CPSF6 insufficiency alters protein levels and causes a developmental syndrome by deregulating APA throughout the transcript. In neonatal humans and zebrafish larvae, CPSF6 insufficiency shifts poly(A) site usage between the 3′UTR and internal sites in a pathway-specific manner. Genes associated with neuronal function undergo mostly intronic APA, reducing their expression, while genes associated with heart and skeletal function mostly undergo 3′UTR APA and are up-regulated. This suggests that, under healthy conditions, cells toggle between internal and 3′UTR APA to modulate protein expression.

Nell'articolo si analizza dal punto di vista della storia politica, come il fascismo italiano ha reagito al verificarsi di disastri naturali, in particolare i terremoti. Vengono quindi presi in considerazione gli strumenti legislativi e tecnici con cui il regime di Mussolini ha affrontato le catastrofi sismiche, nonché le implicazioni politiche conseguenti alla crisi momentanea determinata dall'emergenza, sia in termini di soccorso che di ricostruzione.

Recensione al volume di Michele Cattane, Benedetto Cairoli. Il vessillo della sinistra storica 1825-1889, Carocci, 2020

Il volume ricostruisce la biografia politica dei trenta presidenti del Consiglio (tra i quali Adone Zoni) dei sessantasette governi dell’Italia repubblicana e dei tre governi della transizione. Storici e giuristi intrecciano alle biografie le dimensioni trasversali di tale funzione nevralgica del paese e della macchina sempre più ampia che ha accompagnato il presidente e la presidenza del Consiglio dei ministri, tanto nella sua permanenza nel Palazzo del Viminale, quanto nella nuova residenza di Palazzo Chigi, ed estesasi nei decenni ai palazzi vicini ove sono stati installati dipartimenti, strutture e uffici.

Il saggio intende fornire una riflessione storica di lungo periodo in merito alle politiche ricostruttive messe in atto in Italia dai governi all'indomani di catastrofi naturali.

Recensione al volume di Gianni Silei, I fantasmi della Golden Age. Paura e incertezza nell'immaginario collettivo dell'Europa occidentale (1945-1975), Franco Angeli, 2019

Recensione al volume di Gabriella Gribaudi, La memoria, i traumi, la storia. La guerra e le catastrofi del Novecento, Viella, 2021

L'articolo analizza come le istituzioni pubbliche e le forze politiche in Italia si sono misurate con le emergenze naturali prodotte da fenomeni sismici in epoca repubblicana. Particolare attenzione viene posta agli strumenti e alle strategie con cui i governi hanno affrontato la complessa fase della ricostruzione tenendo conto di una duplice prospettiva: da un lato, i rapporti tra organismi centrali dello Stato e autorità locali, dall'altro, la mai risolta dicotomia tra conservazione e innovazione del contesto urbano e sociale che segna il ritorno alla normalità.

Nel saggio vengono analizzate le modalità e gli strumenti (normativi e pratici) con cui nel corso dell'età liberale le istituzioni pubbliche e le forze politiche hanno reagito in Italia al manifestarsi di eventi catastrofici quali i terremoti.

The RNA-binding protein (RBP) Pumilio1 (PUM1) is associated with two distinct diseases: a late-onset ataxia and a neurodevelopmental syndrome. The ataxia patients retain 75% of normal PUM1 levels, the syndromic patients [~]50%, but this seems inadequate to explain the difference in phenotypes. We hypothesized that mild disease results from dysregulation of PUM1 targets, whereas severe disease involves disruption of PUM1 complexes and deregulation of shared targets. We therefore developed a PUM1 interactome for the murine brain and found that PUM1 shares targets with several RBP interactors (PUM2, FMRP, AGO2, and RBFOX3). PUM1 haploinsufficiency destabilizes these RBPs to varying degrees by brain region and sex, and alters expression of their shared targets, but the milder disease-causing mutation affects only PUM1-specific targets. These data indicate that dosage-sensitive proteins can produce different phenotypes by different mechanisms, and that there may be more intimate cooperation among RBPs than expected. HIGHLIGHTS* RNA-binding proteins can cause disease via their targets or interactors * Interactions among RNA-binding proteins can differ by brain region and sex * A modest reduction in PUM1 levels affects PUM1-specific targets * Greater reductions in PUM1 levels disrupt interactors and shared targets

Il saggio intende fornire un'analisi delle modalità della delegittimazione dell'avversario nelle campagne elettorali italiane d'inizio Novecento, individuandone la natura di strategia volta a porre l’avversario al di fuori dei valori fondativi della comunità politica e di fenomeno sì ricorrente nelle democrazie contemporanee, ma con esiti diversi sulla stabilità dei sistemi politici.

La recensione analizza la raccolta di saggi scritti da Luigi Lacchè nel corso dei suoi studi sulle esperienze costituzionali che hanno scandito tra XIX e XX secolo la storia di Francia, Belgio, Svizzera e Italia

The essay considers the factors and dynamics of the Belgian political crisis of the 1880s. In particular, using the categories of delegitimation, transition, and conflict, it investigates the role played by the monarchy in the clash between Catholics and Liberals during the so-called «School War». Moreover, the essay sheds new light on the constitutional implications of this conflict by placing the Belgian case-study in the more general context of 19th century European liberalism.

Recensione del volume di Bruno Ziglioli dedicato all'emergenza amianto scoppiata a causa della contaminazione di Seveso

Recensione del volume di Valerio Castronovo dedicato alla figura di Giuseppe Venanzio Sella rappresentante di spicco della borghesia imprenditoriale nell'Italia dell'Ottocento.

The genome-wide activity of transcription factors (TFs) on multiple regulatory elements precludes their use as gene-specific regulators. Here we show that ectopic expression of a TF in a cell-specific context can be used to silence the expression of a specific gene as a therapeutic approach to regulate gene expression in human disease. We selected the TF Krüppel-like factor 15 (KLF15) based on its putative ability to recognize a specific DNA sequence motif present in the rhodopsin (RHO) promoter and its lack of expression in terminally differentiated rod photoreceptors (the RHO-expressing cells). Adeno-associated virus (AAV) vector-mediated ectopic expression of KLF15 in rod photoreceptors of pigs enables Rho silencing with limited genome-wide transcriptional perturbations. Suppression of a RHO mutant allele by KLF15 corrects the phenotype of a mouse model of retinitis pigmentosa with no observed toxicity. Cell-specific-context conditioning of TF activity may prove a novel mode for somatic gene-targeted manipulation.

Il saggio ripercorre, attraverso la stampa locale, il contegno delle forze politiche in Valle d'Aosta nei mesi che precedettero l'entrata dell'Italia nella Prima Guerra Mondiale, ponendo particolare attenzione al dibattito sul tema della neutralità sostenuta dal governo allo scoppio del conflitto.

Il saggio intende approfondire quale fu la reazione dell'opinione pubblica in Belgio rispetto al contegno assunto dall'Italia nei dieci mesi compresi tra lo scoppio della Prima guerra mondiale e l'ingresso del nostro paese a fianco dell'Intesa.

Il saggio intende fornire un quadro statistico delle azioni messe in atto dalle forze neutraliste in Italia allo scoppio della Prima guerra mondiale

Transcription factors (TFs) operate by the combined activity of their DNA-binding domains (DBDs) and effector domains (EDs) enabling the coordination of gene expression on a genomic scale. Here we show that in vivo delivery of an engineered DNA-binding protein uncoupled from the repressor domain can produce efficient and gene-specific transcriptional silencing. To interfere with RHODOPSIN (RHO) gain-of-function mutations we engineered the ZF6-DNA-binding protein (ZF6-DB) that targets 20 base pairs (bp) of a RHOcis-regulatory element (CRE) and demonstrate Rho specific transcriptional silencing upon adeno-associated viral (AAV) vector-mediated expression in photoreceptors. The data show that the 20 bp-long genomic DNA sequence is necessary for RHO expression and that photoreceptor delivery of the corresponding cognate synthetic trans-acting factor ZF6-DB without the intrinsic transcriptional repression properties of the canonical ED blocks Rho expression with negligible genome-wide transcript perturbations. The data support DNA-binding-mediated silencing as a novel mode to treat gain-of-function mutations. DOI: http://dx.doi.org/10.7554/eLife.12242.001, eLife digest Proteins called transcription factors bind to sections of DNA known as regulatory elements to activate or deactivate nearby genes. In animals, transcription factors typically have two sections: a “DNA-binding domain” that attaches to DNA, and an “effector domain” that is responsible for interacting with other proteins to regulate the gene’s expression. Rhodopsin is a gene that encodes the instructions needed to make a light-sensitive protein in the eyes of humans and other animals. Botta et al. have now used this gene as an example to investigate whether proteins that contain a DNA-binding domain – but not an effector domain – can repress gene expression. The experiments show that only a small section of the regulatory elements in the human Rhodopsin gene is actually required for the gene to be expressed. Botta et al. designed an artificial protein – referred to as ZF6-DB – that is able to bind to this section of DNA. The binding of ZF6-DB to this short DNA section was sufficient to switch off a Rhodopsin gene in living pig cells, and, unlike conventional transcription factors, seemed to have minimal impact other genes. Next, Botta et al. used a virus to insert both the gene that encodes ZF6-DB and a normal copy of Rhodopsin into pigs. In these animals, ZF6-DB switched off the existing copy of Rhodopsin, but not the inserted copy so the cells produced a working form of the light-sensitive protein. Further experiments were carried out in mice that have both a faulty version and a normal copy of the Rhodopsin gene. ZF6-DB switched off the faulty Rhodopsin gene, which allowed the normal Rhodopsin gene to work without any interference from the faulty copy. Mutations in Rhodopsin can cause an eye disease that leads to severe loss of vision in humans. These new findings could now guide future efforts to develop treatments for people with this condition. It will also be important to investigate how ZF6-DB binds to the regulatory elements in the Rhodopsin gene and whether a similar strategy could be used to alter the expression of other genes. DOI: http://dx.doi.org/10.7554/eLife.12242.002

Silencing and replacement strategy is a promising strategy to overcome mutational heterogeneity of genetic defects. In autosomal dominant retinitis pigmentosa (adRP) approximately 200 different mutations have been associated with rhodopsin gene (RHO). We recently described a system based on the delivery of a synthetic transcriptional repressor (DNA-binding-mediated silencing; ZF6-DBD), which upon Adeno-associated viral (AAV) vector delivery to the porcine retina, demonstrated selective somatic ablation of RHO in mutation independent manner, thus silencing both the wild-type and mutated alleles. To determine the actual therapeutic impact of DNA-binding-mediated silencing, we carried out the silencing-replacement strategy by coupling ZF6-DBD with RHO replacement (human RHO, hRHO CDS). To warrant simultaneous photoreceptor transduction of both ZF6-DBD and hRHO, we enclosed two expression cassettes into a single vector (DNA-binding repression and replacement, DBR-R). To generate the construct we evaluated key variables for balanced simultaneous RHO repression and replacement, which are the vector dose and promoter strength. To ensure high and rod-specific hRHO replacement, we opted for a high vector dose and the strength of the GNAT1 promoter elements. To decrease ZF6-DB expression levels at high vector dose, while keeping rod-specificity, we both shortened the human RHO promoter and deleted the 5’ sequence of the ZF6-DB target. We used 1×1012 gc of vector of DBR-R (AAV8-RHOΔ-ZF6-DB-GNAT1-hRHO) to administer to porcine retina. Administration of the DBR-R vector resulted in concomitant rod-specific transcriptional repression of the porcine Rho (35%) and in balanced replacement with the exogenous hRHO (45%), as assessed by transcripts levels, protein expression and integrity of photoreceptor outer segments. The data support pre-clinical development including a dose-response and long-term efficacy and toxicity.

Transcription factors (TFs) function by the combined activity of their DNA-binding domains (DBDs) and effector domains (EDs). Here we show that in vivo delivery of an engineered DNA-binding protein uncoupled from the repressor domain entails complete and gene-specific transcriptional silencing. To silence RHODOPSIN (RHO) gain-of-function mutations, we engineered a synthetic DNA-binding protein lacking canonical repressor domains and targeted to the regulatory region of the RHO gene. AAV-mediate retinal delivery at a low dose (AAV2/8-CMV-ZF6-DBD, 1×10e10 vector genomes, vg) in the porcine retina resulted in selective transcriptional silencing of RHO expression. The rod photoreceptors (the RHO expressing cells) transduced cells when isolated by FACS-sorting showed the remarkable 90% RHO transcriptional repression. To evaluate genome-wide transcriptional specificity, we analyzed the porcine retina transcriptome by RNA sequencing (RNA-Seq). The differentially expressed genes (DEGs) analysis showed that only 19 genes were perturbed. In this study, we describe a system based on a synthetic DNA binding protein enabling targeted transcriptional silencing of the RHO gene by in vivo gene transfer. The high rate of transcriptional silencing occurring in transduced cells supports applications of this regulatory genomic interference with a synthetic trans-acting factor for diseases requiring gene silencing in a large number of affected cells, including for instance a number of neurodegeneration disorders. The result support a novel mode of gene targeted silencing with a DNA-binding protein lacking intrinsic activity.

Silencing and replacement strategy is a promising approach to overcome mutational heterogeneity of genetic defects. In autosomal dominant retinitis pigmentosa (adRP) due to rhodopsin gene (RHO) approximately 200 different mutations have been described, posing a challenge for the design of effective therapeutics.We designed a silencing and replacement strategy based on transcriptional silencing through an artificial zinc finger DNA-binding protein lacking effector domains (ZF6DBD), and tested both efficacy and safety in two animal models.In a murine model of adRP, we show that AAV-mediate retinal delivery (AAV2/8-CMV-ZF6-DBD) is associated with selective transcriptional silencing of the human mutated allele resulting in morphological and functional (Electroretinography, ERG a-wave and b-wave responses) rescue. We then tested the effect of transcriptional silencing in the porcine large pre-clinical model. Delivery of a low dose (AAV2/8-CMV-ZF6-DBD, 1×10e10 vector genomes, vg) of the ZF6 transcriptional silencer to the porcine retina resulted in robust transcriptional silencing of the endogenous porcine RHO transcript. Cell sorting of transduced photoreceptors showed an almost complete RHO transcriptional silencing effect (90% RHO transcriptional repression), underscoring the potency of the system. To determine the safety of the zinc-finger silencer we performed extensive RNA-seq analysis on treated and control retinae. The data sets generated demonstrate selective RHO gene transcriptional repression and a remarkably low number of differential expressed genes (DEGs), supporting specificity and thus, safety. The co-administration to the porcine retina of the AAV-ZF6 silencer (AAV2/8-CMV-ZF6-DBD) and the AAV-RHO replacement (5×10e11 vg, AAV2/8-GNAT1-HumanRHO) constructs resulted in a balanced silencing and replacement effect. This data support the use of zinc-finger based RHO transcriptional silencing for the development of a clinical trial for adRP patients.