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Microplastics represent a threat due to their ability to enter the food chain, with harmful consequences for living organisms. The riskiness of these particles is also linked to the release of other contaminants, such as heavy metals. Solute Carriers (SLCs) represent eminent examples of first-level targets of heavy metals due to their localization on the cell surface. Putative targets of heavy metals are the organic cation transporters that form a sub-clade of the SLC22 family. Besides the physiological role in the absorption/release of endogenous organic cations, these transporters are crucial in drug disposition and their interaction with xenobiotics. In this work, the human SLC22A4, commonly known as OCTN1, was used as a benchmark to test interactions with heavy metals released by microplastics, exploiting the proteoliposome tool. The potency of metals to interfere with the OCTN1 function has been evaluated by measuring IC50 values calculated in the micromolar range. The molecular mechanism of interaction has been defined using site-directed mutagenesis and computational analyses. Finally, some chemical and physiological thiol-reacting compounds show the capacity to rescue the metal-inhibited OCTN1 function. The conclusions drawn on OCTN1 can be extended to other members of the SLC22 family and orthologous transporters in fish.

Health risks are often overlooked when the short-term consequences are not immediately apparent. During restoration work, cleaning actions can generate particles that pose health risks to workers through inhalation. This is particularly true in the case of asbestos fibres that might be spread out from the laser cleaning of buildings or heritage artifacts made of stone, such as serpentinite and other ultramafic rocks, that have a high probability of containing asbestos (e.g., chrysotile, tremolite asbestos, actinolite asbestos). To show workers the importance of wearing proper protection to prevent health injuries, several serpentinite samples, ascertained to contain asbestos minerals by specific investigations, have been laser ablated using ad hoc modified equipment in order to collocate a HEPA filter prone to collect all dust emitted during ablation. The powder deposited on the surface of these filters after laser ablation was analyzed, by Powder X -ray Diffraction (PXRD), Transmission Electron Microscopy combined with Energy Dispersive Spectroscopy (TEM/EDS), micro-Raman spectroscopy (μ-Raman) and Fourier-transform infrared spectroscopy (FTIR/ATR). The results confirmed the presence of asbestos fibres during the laser ablation of rocks containing Naturally Occurring Asbestos (NOA), emphasizing the importance of wearing appropriate personal protective equipment (PPE) during these procedures. Noteworthy, approximately 33 % of the analyzed fibres met the WHO criteria in size for respirable fibres. Furthermore, through our experiments, we also demonstrated that using tools that integrate filters into working tools would definitively further decrease the risk of fibres inhalation to workers., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Most of the artworks stored in museums are often kept in inappropriate climatic and environmental conditions that facilitate the formation and growth of microorganisms, such as fungi, which are responsible for many types of biodegradation phenomena. To mitigate and prevent these deteriorative processes, functionalized packaging materials can be used for the storage and handling of artworks. The aim of this study was to develop a potential anti-biodeterioration coating suitable for packaging purposes. TiO 2 and ZnO doped with different amounts of Ag (0.5 wt%, 1 wt%, and 3 wt%) were synthesized and dispersed in polyvinyl alcohol (PVA) and acrylic resin (Paraloid B72), then applied on different types of packaging materials (cellulose and the high-density spunbound polyethylene fiber Tyvek ® , materials that are frequently used as packaging in museums). Analytical investigations (SEM/EDS, Raman, FTIR, and XRD) were employed to assess dispersion on the packaging material. Furthermore, resistance against biodeteriogens was assessed using Cladosporium sp., a bioluminometer, to define the biocidal efficacy.

The main conservation problem of p-PVC artworks is phthalate-based plasticizer migration. Phthalate migration from the bulk to the surface of the materials leads to the formation of a glossy and oily film on the outer layers, ultimately reducing the flexibility of the material. This study aimed to develop a removable coating for the preservation of contemporary artworks and design objects made of plasticized polyvinyl chloride (p-PVC). Several coatings incorporating chitosan, collagen, and cellulose ethers were assessed as potential barriers to inhibiting plasticizer migration. Analytical techniques including optical microscopy (OM), ultraviolet/visible/near-infrared spectroscopy (UV/Vis/NIR), Fourier transform infrared spectroscopy with attenuated total reflection (FTIR-ATR), and scanning electron microscopy (SEM) were utilized to evaluate the optical and chemical stability of selected coating formulations applied to laboratory p-PVC sheet specimens. Subsequently, formulations were tested on a real tangible example of a design object, ©Barbie doll, characterized by the prevalent issue of plasticizer migration. Furthermore, the results obtained with the tested formulations were evaluated by a group of conservators using a tailored survey. Finally, a suitable coating formulation capable of safeguarding plastic substrates was suggested.

Modern society and industrial development rely heavily on the availability of freshwater and minerals. Seawater reverse osmosis (SWRO) has been widely adopted for freshwater supply, although many questions have arisen about its environmental sustainability owing to the disposal of hypersaline rejected solutions (brine). This scenario has accelerated significant developments towards the hybridization of SWRO with membrane distillation-crystallization (MD-MCr), which can extract water and minerals from spent brine. Nevertheless, the substantial specific energy consumption associated with MD-MCr remains a significant limitation. In this work, energy harvesting was secured from renewables by hotspots embodied in the membranes, implementing the revolutionary approach of brine mining via photothermal membrane crystallization (PhMCr). This method employs self-heating nanostructured interfaces under solar radiation to enhance water evaporation, creating a carefully controlled supersaturated environment responsible for the extraction of minerals. Photothermal mixed matrix photothermal membranes (MMMs) were developed by incorporating graphene oxide (GO) or carbon black (CB) into polyvinylidene fluoride (PVDF) solubilized in an eco-friendly solvent (i.e., triethyl phosphate (TEP)). MMMs were prepared using non-solvent-induced phase separation (NIPS). The effect of GO or GB on the morphology of MMMs and the photothermal behavior was examined. Light-to-heat conversion was used in PhMCr experiments to facilitate the evaporation of water from the SWRO brine to supersaturation, leading to sodium chloride (NaCl) nucleation and crystallization. Overall, the results indicate exciting perspectives of PhMCr in brine valorization for a sustainable desalination industry.

Over the centuries, humans have developed different systems to protect surfaces from the influence of environmental factors. Protective paints are the most used ones. They have undergone considerable development over the years, especially at the turn of the 19th and 20th centuries. Indeed, between the two centuries, new binders and pigments have been introduced in the constituent materials of paints. The years in which these compounds have been introduced and spread in the paint market allow them to be defined as markers for the dating of paints and painted artifacts. The present work is focused on the study of the paints of two vehicles of the Frankfurt Museum of Communication, i.e., a carriage and a cart, that was designed for the German Postal and Telecommunications Service roughly between 1880 and 1920. The characterization of the paints was performed through in situ non-invasive techniques, i.e., portable optical microscopy and multispectral imaging, and laboratory non-destructive techniques, i.e., FT-IR ATR spectroscopy and SEM-EDS. The analytical investigation and the comparison with the data reported in the literature allowed us to determine the historicity of the paints, which are all dated before the 1950s.

In the present study, a diagnostic approach was used to analyze the wall painting in the apse of the Sotterra church at Paola, in the province of Cosenza, Italy. The Sotterra church is nowadays located 6 m under the ground level. The presbytery area houses valuable pictorial evidence attributable to different phases. The oldest painting, visible in the apse area, dates back to the 11th-12th centuries, and it represents the subject of the present study, while the later decorations are placed in a chronological range from the 14th to the 15th centuries. Due to the peculiar environmental conditions, the conservation of subterranean sites represents a debated issue and must be properly investigated. For this reason, in this research, noninvasive analysis and laboratory-based methods were planned to obtain information about both the composition of original materials and the mechanisms and causes of alterations affecting the wall painting in the apse. Simultaneously, an environmental monitoring campaign of the indoor climate for the duration of nine months was conducted. The results highlighted the use of natural mineral pigments such as ochres and earths. The analysis of the painting stratigraphy revealed that the mural painting consists of two plaster layers characterized by lime-based binder. Moreover, the presence of a high amount of calcium sulfate has been discovered; this latter result, combined with the monitoring of the microclimate, allowed for the establishment of the crystallization and the condensation risks which occur on the investigated surfaces.

Mattel's Barbie dolls are the most famous and iconic dolls since 1959. Today, they are being collected by individuals and often conserved in museum environments due to their cultural and historical significance reflecting everyday life and historical events. However, just like most museum objects made of plastics, both historical and more recent Barbies show evident degradation phenomena. Firstly, Barbies were made of plasticized polyvinyl chloride (PVC), affected by the migration of additives-mostly the plasticizers-from the bulk phase to the outermost layer, appearing as a tacky and glossy exudate. Over the years, Barbies' polymeric constituents were replaced with more stable ones, whose additives migration is limited compared to PVC, even though still occurring. Multispectral photography in visible (VIS) and ultraviolet (UV) light, microscopical observations in VIS and UV light, and Fourier Transform Infrared spectroscopy in the Attenuated Total Reflectance mode (FT-IR ATR) were performed to characterize the constituent materials of 15 Barbies produced between 1959 and 1976, bridging the information gap on their processing over the years. The micro-invasive multi-analytical approach also allowed for the characterization of the degradation products, permitting the reference of the exudated compound to the specific bulk polymer.

Biodegradation is among the most common issues affecting Cultural Heritage stone materials in outdoor environments. In recent years, the application of chemical agents with biocidal activity has been the most usual practice when dealing with biofilm removal. In outdoor environments, the use of these biocides is not effective enough, since the materials are constantly exposed to environmental agents and atmospheric pollutants. Thus, it becomes necessary to protect the surface of Cultural Heritage works with antimicrobial coatings to either prevent or at least limit future colonization. In this study, innovative biocides-both natural and synthetic-were applied on a Roman mosaic located in the Archaeological Park of Ostia Antica to compare their effectiveness in removing the biological degradation affecting it. In addition, an antimicrobial coating called "SI-QUAT" was applied and analyzed in situ. SI-QUAT has recently entered the market for its prevention activity against biocolonization. The biocidal activity of these products was tested and monitored using different analytical portable instruments, such as the multispectral system, the spectrocolorimeter, and the bioluminometer. The analyses showed that promising results can be obtained using the combination of the biocide and the protective effect of Preventol ® RI50 and SI-QUAT.

In this study, the investigation of the oil painting on canvas I Tesori del Mare made by Plinio Nomellini in 1901 is presented. The aim of the research was threefold: the examination of the state of conservation in view of the restoration treatment, together with the identification of the causes of degradation and the study of the artistic technique. During the years, the artwork underwent several cleaning and fixing interventions, resulting in a patchy appearance of the surface. Nevertheless, the presence of consistent liftings persists, while the protective coating shows uneven chromatic alteration, both requiring further analysis. Multispectral imaging allowed for better visualization of the figuration's structure and the restored areas. The combined use of Raman spectroscopy, Fourier Transform Infrared spectroscopy in the Attenuated Total Reflection mode (FT-IR ATR), and Scanning Electron Microscopy coupled with an Energy Dispersive Spectroscopy (SEM/EDS) enabled better understanding of the stratigraphy through the identification of some pigments, the binder, and the aged varnish layer on the top. SEM/EDS highlighted the presence of zinc in both the ground layer and the paint layers. Furthermore, FT-IR ATR spectroscopy showed peaks related to metal soaps such as zinc stearate, which are known to cause severe delamination of the paint layers, explaining the recurring lifting issues. Eventually, the varnish layer was found to be acrylic resin, presumably mixed with varnishes applied in past restoration treatments.

Biodeterioration is an increasingly widespread process of degradation in the context of the conservation of cultural heritage, which involves a combination of physical and chemical damages together with an aesthetic alteration of materials. For biological damage on monuments caused by pathogens, macro- and microorganisms, chemical treatments are generally used, most of the time dangerous for the environment and for the operator. In this context, new eco-friendly products represent necessary tools for the treatment of biologically deteriorated stone surfaces and represent a new challenge in the field of restoration and conservation of materials of cultural interest. A relatively new class of unconventional green solvents are deep eutectic solvents (DESs), which have peculiar chemical-physical characteristics such as being non-toxic, ecological, biodegradable, non-flammable, and stable in the presence of water. Furthermore, many DESs known in the literature have also been shown to have a biocidal action. All these characteristics make DESs very advantageous and safe, and they could be used as biocidal agents for the treatment of biodegraded surfaces of cultural heritage, being non-toxic for the environment and for the operator. So far, they are used in various fields, but they still represent a novel frontier in the cultural heritage sector. The present research aims at testing five different DESs for the first time in cultural heritage. In particular, DESs are applied to a mosaic located in the Ostia Antica Archaeological Park (Rome), and their efficiency is compared with a biocide product currently used in the restoration field, namely, Preventol RI50, through luminescence, bio-luminometry, and spectrocolorimetry analysis. The preliminary results achieved show the different behaviors of each DESs, highlighting the possibility of employing them in the field of cultural heritage. Further studies have been planned, some of which are already underway, to investigate the properties of DESs and indicate any improvements to make them more effective, both as solvents and as biocides, and easy to apply to various types of materials. The results obtained from this first study are very promising for the use of DES as a new green strategy for cleaning and conservation treatments of materials in the field of cultural heritage.

A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of the viral titer. We show that Cas13d-mediated coronavirus inhibition is dependent on the crRNA cellular spatial colocalization with Cas13d and target viral RNA. Cas13d can significantly enhance the therapeutic effects of diverse small molecule drugs against coronaviruses for prophylaxis or treatment purposes, and the best combination reduced viral titer by over four orders of magnitude. Using lipid nanoparticle-mediated RNA delivery, we demonstrate that the Cas13d system can effectively treat infection from multiple variants of coronavirus, including Omicron SARS-CoV-2, in human primary airway epithelium air-liquid interface (ALI) cultures. Our study establishes CRISPR-Cas13 as a BSA which is highly complementary to existing vaccination and antiviral treatment strategies., (© 2022. The Author(s).)

A multi-analytical approach was employed to study wall paintings located in the Sotterra church at Paola, in the province of Cosenza, Italy. The site is an underground church (hence the name of Sotterra, which means "under the earth") rediscovered in the second half of the 19th century, during the building works of the Madonna del Carmine church on the same area. This underground church preserves valuable mural paintings having different styles. The construction's dating and overlapped modifications made until the site was abandoned is also debated. A wall painting, depicting "The Virgin" as part of the "Annunciation and the Archangel Gabriel" present on the opposite side of the apse, was selected and investigated using both in situ and laboratory-based analysis. Preliminarily, the non-destructive investigations involved several analytical techniques (IR imaging, UV-Induced Visible Fluorescence, and X-ray Fluorescence analyses) that provided mapping and characterization of pictorial layers and first data about deterioration phenomena. On the basis of this information, a more in-depth study was conducted on micro-fragments aimed at characterizing the stratigraphy and to identify the artist's technique. Cross-sections were analysed using polarized optical microscopy and electron scanning microscopy coupled with energy-dispersive X-ray spectroscopy to obtain morphological and chemical information on the selected pictorial micro-fragments of the wall painting. The results allowed to characterize the pigments and provide better readability of the whole figure, revealing details that are not visible to the naked eye, important for future historical-artistic and conservative studies. The results represent the first step of a systematic archaeometric research aimed at supporting the ongoing historical-stylistic studies to distinguish the different building phases hypothesized for this religious site which remained buried for three centuries.

This research has focused on the assessment of the compositional features and mechanical and antifouling performances of two different mortars formulated for an underwater setting, and which contain Mg(OH) 2 as an antifouling agent. Regarding the mechanical characterization, the uniaxial compressive strength and flexural strength were measured. The composition of the materials was explored by differential thermal/thermogravimetric analysis (DTA-TG), X-ray diffraction analysis (XRPD), and scanning electronic microscopy (SEM) coupled with EDS microanalysis. The assessment of the biological colonization was evaluated with colorimetric analysis and image analysis. The results suggest that both mortars have good mechanical resistance once set underwater. Moreover, the adding of Mg(OH) 2 improves the resistance toward biofouling; this was observed both in laboratory and sea-exposed specimens.

Bio-colonization is a dynamic and multiphasic process headed by microorganisms. Conventional treatments to process affected stone materials include chemical biocides, whose formulations are mainly composed of quaternary ammonium salts(QAs), reported to be toxic for human health, dangerous for the environment, and not biodegradable. Accordingly, novel green and eco-friendly products are a promising alternative to treat stone materials deteriorated by microorganism colonization. In this study, the efficacy of pure essential oils (EOs) and a mix of EOs was assessed in situ and compared to a conventional biocide based on QAs, and two commercially green products based on EOs, which were taken as references, through application on a mosaic located at the Archaeological Park of Ostia Antica (Rome). The EO biocide efficacy was analyzed by ultraviolet induced luminescence, spectro-colorimetry and bio-luminometry analyses while the possibility of their permanence on simulated substrate was studied by FTIR spectroscopy. It was observed by FTIR analysis, that EOs considered volatile can leave a residue after the application; typical fingerprint bands at about 2926, 1510, and 1455 cm -1 were recorded in the EO spectra. Every tested oil was confirmed to have a biocide action although minimal in relation to the most conventional products based on QAs. The synergy of the essential oils revealed positive results, showing a stronger biocide efficacy. Further investigation should be carried out to develop the method of application and study of essential oils on cultural heritage.

Urban art is a form of artistic visual expression and communication that is created in the street and generally in the public dimension of urban spaces. Often these kinds of artworks are in outdoor environments, and they usually suffer from atmospheric weathering and anthropic vandalism. Recently, several strategies have been used to limit or remove the effects of such vandalism. Currently, the use of quartz paints is growing among artists; such paints after setting are more porous and rough on the surface with respect to regular paints. The aim of the study is to assess the performance of anti-graffiti coatings on quartz artworks paints. Two anti-graffiti products were chosen, and their behaviors were assessed in the laboratory by means of contact angle measurement, water capillary test, colorimetric analysis, and optical and electron microscopy. Results showed good water repellence efficacy of the tested products, demonstrating that they are suitable for the protection of urban art, but at least two applications on the surface are needed to achieve good performance.

Antarctica represents a unique natural laboratory for ecotoxicological studies as it is characterized by low internal pollutants emissions but high external contamination levels. Indeed, warm temperatures promote pollutant evaporation (low latitudes), while cool temperatures (high latitudes) promote its deposition from the atmosphere on land/water. Metals are the most important pollutants in ecosystems and represent a serious and global threat to aquatic and terrestrial organisms. Since 2000, the risks posed by metals have led many States to ratify protocols aimed at reducing their emissions. Endemic Antarctic organisms represent excellent bioindicators in order to evaluate the efficacy of global measures adopted to mitigate pollutants release into the environment. In this study (supported by PNRA18-00133), we estimated the metals contamination levels and the metallothionein-1 expression in liver samples of two Antarctic fish species, the icefish Chionodraco hamatus and the red-blooded Trematomus bernacchii , collected in the same area during 2002 and 2014. The chosen area is located in the Ross Sea, a unique area as it is also isolated from the rest of the Southern Ocean. The analysis of contamination trends throughout this period showed, in both species, a significant increase over time of metals bioaccumulation and metallothionein-1 expression. In addition, our result clearly indicated that the detoxifying ability of the two organisms analyzed greatly differs, probably due to haemoglobin presence/absence. Our work represents an important early step to obtain valuable information in conservation strategies for both Antarctic and non-Antarctic ecosystems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Marrone, La Russa, Brunelli, Santovito, La Russa, Barca and Pellegrino.)

Bloodstains found at crime scenes represent a crucial source of information for investigative purposes. However, in forensic practice, no technique is currently used to estimate the time from deposition of bloodstains. This preliminary study focuses on the age estimation of bloodstains by exploiting the color variations over time due to the oxidation of the blood. For this purpose, we used a colorimetric methodology in order to easily obtain objective, univocal and reproducible results. We developed two bloodstain age prediction algorithms: a short-term and a long-term useful model for the first 24h and 60 days, respectively. Both models showed high levels of classification accuracy, particularly for the long-term model. Although a small-scale study, these results improve the potential application of colorimetric analysis in the time-line reconstruction of violent criminal events.

Plastic is everywhere-increasing evidence suggests that plastic pollution is ubiquitous and persistent in ecosystems worldwide. Microplastic pollution in marine environments is particularly insidious, as small fragmentation can increase interaction with biota and food chain access. Of particular concern is the Mediterranean Sea, which has become a large area of accumulation of plastic debris, including microplastics, whose polymeric composition is still largely unknown. In this study, we analyzed the polymeric composition, particle size distribution, shape, and color of small plastic particles (ranging from 50 to 5000 µm) collected from the sea surface in six stations at the center of the Mediterranean Sea. We also described, for the first time, the different distribution of microplastics from coastal areas up to 12 nautical miles offshore. The microplastic density was 0.13 ± 0.19 particles/m 2 , with a marked prevalence of smaller particles (73% < 3 mm) and a peak between 1 and 2 mm (34.74%). Microplastics composition analysis showed that the most abundant material was polyethylene (69%), followed by polypropylene (24%). Moreover, we reported a comparison of the two Calabrian coasts providing the first characterization of a great difference in microplastic concentration between the Tyrrhenian and Ionian sides (87% vs. 13%, respectively), probably due to the complex marine and atmospheric circulation, which make the Tyrrhenian side an area of accumulation of materials originating even from faraway places. We demonstrate, for the first time, a great difference in microplastic concentration between Tyrrhenian and Ionian Calabrian coasts, providing a full characterization and highlighting that microplastic pollution is affected by both local release and hydrography of the areas.

The coronavirus disease 2019 (COVID-19) pandemic, caused by the SARS-CoV-2 virus, has highlighted the need for antiviral approaches that can target emerging viruses with no effective vaccines or pharmaceuticals. Here, we demonstrate a CRISPR-Cas13-based strategy, PAC-MAN (prophylactic antiviral CRISPR in human cells), for viral inhibition that can effectively degrade RNA from SARS-CoV-2 sequences and live influenza A virus (IAV) in human lung epithelial cells. We designed and screened CRISPR RNAs (crRNAs) targeting conserved viral regions and identified functional crRNAs targeting SARS-CoV-2. This approach effectively reduced H1N1 IAV load in respiratory epithelial cells. Our bioinformatic analysis showed that a group of only six crRNAs can target more than 90% of all coronaviruses. With the development of a safe and effective system for respiratory tract delivery, PAC-MAN has the potential to become an important pan-coronavirus inhibition strategy., Competing Interests: Declaration of Interests The authors have filed provisional patents via Stanford University related to this work., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The Tomb of the Diver has been subject for many decades of fierce debate among archaeologists and classicists. Since its discovery in 1968, some scholars have considered it a unique example of the lost tradition of Greek painting, others have emphasized Etruscan or Italic parallels. More recently, a possible local production has been suggested. With the aim of trying to solve the archaeological question, an archaeometric comparison among this well-known artwork and several frescoed tombs coming from Hellenistic and Lucan necropolis was carried out. The multi-analytical study was focused on the identification of peculiar features of executive techniques and raw materials since the first period of the archaeological site. The analytical investigation has been preliminary based on a non-destructive approach, performed in-situ by portable equipment including imaging diagnostics and compositional spectroscopic techniques for identifying pigments and the conservation state of original painted surface; subsequently, a further deepening by using destructive techniques was performed in-lab for the mortar-based supports characterization. Archaeometric study suggested that technological choices slightly changed in a time span of about two centuries, highlighting important markers that allow clustering the contemporary artistic productions. Moreover, a comparison with mortars from temples decorations was provided to better understand the whole artistic context. The archaeometric data showed that the Tomb of the Diver could be traced back to a local artisanal tradition and therefore is neither Etruscan nor Greek, but the first and foremost an expression of the local elite culture of Paestum., Competing Interests: The authors have declared that no competing interests exist. The commercial affiliations do not alter our adherence to PLOS ONE policies on sharing data and materials.

CRISPR-Cas9 technology has rapidly changed the landscape for how biologists and bioengineers study and manipulate the genome. Derived from the bacterial adaptive immune system, CRISPR-Cas9 has been coopted and repurposed for a variety of new functions, including the activation or repression of gene expression (termed CRISPRa or CRISPRi, respectively). This represents an exciting alternative to previously used repression or activation technologies such as RNA interference (RNAi) or the use of gene overexpression vectors. We have only just begun exploring the possibilities that CRISPR technology offers for gene regulation and the control of cell identity and behavior. In this review, we describe the recent advances of CRISPR-Cas9 technology for gene regulation and outline advantages and disadvantages of CRISPRa and CRISPRi (CRISPRa/i) relative to alternative technologies., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)