Antisense oligonucleotide (ASO)-mediated gene silencing has broad applications, spanning from biomedicine to agriculture, involving molecular biology, synthetic biology, and genetic manipulation. ...This research harnessed nanotechnology to augment ASO-mediated gene silencing, introducing a remotely switchable gene expression system for precise temporal control. We targeted lipid biosynthesis and accumulation enhancement in the photosynthetic eukaryote Chlamydomonas reinhardtii. Gold nanoparticles (AuNPs) transported double-stranded DNA (dsDNA), forming dsDNA–AuNP complexes. These complexes comprised 3′-thiolated sense strands attached to AuNPs and fluorescent antisense oligonucleotides. To avoid harmful laser effects on cells, we adopted a light-emitting diode (LED). Confocal microscopy confirmed dsDNA–AuNP internalization in C. reinhardtii. LED-triggered antisense release led to an 83% decrease in Citrate Synthase 2 (CIS 2) expression. Thiolated sense strand attachment postillumination inhibited antisense reannealing, enhancing gene silencing. This led to significant lipid body accumulation in cells, verified through fluorometric and fluorescence microscopy. This union of nanotechnology and ASO-mediated silencing provides gene regulation opportunities across sectors like biomedicine and agriculture. The system’s remote switching capability underscores its potential in synthetic biology and genetic engineering. Our findings substantiate the utility of this approach for enhancing lipid biosynthesis in C. reinhardtii but also underscores its broader applicability to other organisms, fostering the development of novel solutions for pressing global challenges in energy, agriculture, and healthcare.
Low-cost substrates are an exciting alternative for bioprocesses; however, their complexity can affect microorganism metabolism with non-desirable outcomes. This work evaluated banana peel extract ...(BPE) as a growth medium compared to commercial Yeast-Malt (YM) broth in the native and non-conventional yeast Rhodotorula mucilaginosa UANL-001L. The production of carotenoids, fatty acids, and exopolysaccharides (EPS) was also analyzed. Biomass concentration (3.9 g/L) and growth rate (0.069 g/h) of Rhodotorula mucilaginosa UANL-001L were obtained at 200 g/L of BPE. Yields per gram of dry biomass for carotenoids (317 µg/g) and fatty acids (0.55 g/g) showed the best results in 150 g/L of BPE, while 298 µg/g and 0.46 mg/g, respectively, were obtained in the YM broth. The highest yield of EPS was observed in 50 g/L of BPE, a two-fold increase (160.1 mg/g) compared to the YM broth (76.3 mg/g). The fatty acid characterization showed that 100 g/L of BPE produced 400% more unsaturated compounds (e.g., oleic and ricinoleic acid) than the YM broth. Altogether, these results indicate that BPE is a suitable medium for producing high-value products with potential industrial applications.
The use of fossil fuels has been strongly related to critical problems currently affecting society, such as: global warming, global greenhouse effects and pollution. These problems have affected the ...homeostasis of living organisms worldwide at an alarming rate. Due to this, it is imperative to look for alternatives to the use of fossil fuels and one of the relevant substitutes are biofuels. There are different types of biofuels (categories and generations) that have been previously explored, but recently, the use of microalgae has been strongly considered for the production of biofuels since they present a series of advantages over other biofuel production sources: (a) they don't need arable land to grow and therefore do not compete with food crops (like biofuels produced from corn, sugar cane and other plants) and; (b) they exhibit rapid biomass production containing high oil contents, at least 15 to 20 times higher than land based oleaginous crops. Hence, these unicellular photosynthetic microorganisms have received great attention from researches to use them in the large-scale production of biofuels. However, one disadvantage of using microalgae is the high economic cost due to the low-yields of lipid content in the microalgae biomass. Thus, development of different methods to enhance microalgae biomass, as well as lipid content in the microalgae cells, would lead to the development of a sustainable low-cost process to produce biofuels. Within the last 10 years, many studies have reported different methods and strategies to induce lipid production to obtain higher lipid accumulation in the biomass of microalgae cells; however, there is not a comprehensive review in the literature that highlights, compares and discusses these strategies. Here, we review these strategies which include modulating light intensity in cultures, controlling and varying CO
levels and temperature, inducing nutrient starvation in the culture, the implementation of stress by incorporating heavy metal or inducing a high salinity condition, and the use of metabolic and genetic engineering techniques coupled with nanotechnology.
Since the discovery of antibiotics, humanity has been able to cope with the battle against bacterial infections. However, the inappropriate use of antibiotics, the lack of innovation in therapeutic ...agents, and other factors have allowed the emergence of new bacterial strains resistant to multiple antibiotic treatments, causing a crisis in the health sector. Furthermore, the World Health Organization has listed a series of pathogens (ESKAPE group) that have acquired new and varied resistance to different antibiotics families. Therefore, the scientific community has prioritized designing and developing novel treatments to combat these ESKAPE pathogens and other emergent multidrug-resistant bacteria. One of the solutions is the use of combinatorial therapies. Combinatorial therapies seek to enhance the effects of individual treatments at lower doses, bringing the advantage of being, in most cases, much less harmful to patients. Among the new developments in combinatorial therapies, nanomaterials have gained significant interest. Some of the most promising nanotherapeutics include polymers, inorganic nanoparticles, and antimicrobial peptides due to their bactericidal and nanocarrier properties. Therefore, this review focuses on discussing the state-of-the-art of the most significant advances and concludes with a perspective on the future developments of nanotherapeutic combinatorial treatments that target bacterial infections.
Gold nanoparticles (AuNPs) can be found in different shapes and sizes, which determine their chemical and physical characteristics. Physical and chemical properties of metallic NPs can be tuned by ...changing their shape, size, and surface chemistry; therefore, this has led to their use in a wide variety of applications in many industrial and academic sectors. One of the features of metallic NPs is their ability to act as optothermal energy converters, where they absorb light at a specific wavelength and heat up their local nanosurfaces. This feature has been used in many applications where metallic NPs get coupled with thermally responsive systems to trigger an optical response. In this study, we synthesized AuNPs that are spherical in shape with an average diameter of 20.07 nm. This work assessed simultaneously theoretical and experimental techniques to evaluate the different factors that affect heat generation at the surface of AuNPs when exposed to a specific light wavelength. The results indicated that laser power, concentration of AuNPs, time × laser power interaction, and time illumination, were the most important factors that contributed to the temperature change exhibited in the AuNPs solution. We report a regression model that allows predicting heat generation and temperature changes with residual standard errors of less than 4%. These results are highly relevant in the future design and development of applications where metallic NPs are incorporated into systems to induce a temperature change triggered by light exposure.
A declining pipeline of clinically useful antibiotics has made it imperative to develop more effective antimicrobial therapies, particularly against difficult-to-treat Gram-negative pathogens. Silver ...has been used as an antimicrobial since antiquity, yet its mechanism of action remains unclear. We show that silver disrupts multiple bacterial cellular processes, including disulfide bond formation, metabolism, and iron homeostasis. These changes lead to increased production of reactive oxygen species and increased membrane permeability of Gram-negative bacteria that can potentiate the activity of a broad range of antibiotics against Gram-negative bacteria in different metabolic states, as well as restore antibiotic susceptibility to a resistant bacterial strain. We show both in vitro and in a mouse model of urinary tract infection that the ability of silver to induce oxidative stress can be harnessed to potentiate antibiotic activity. Additionally, we demonstrate in vitro and in two different mouse models of peritonitis that silver sensitizes Gram-negative bacteria to the Gram-positive-specific antibiotic vancomycin, thereby expanding the antibacterial spectrum of this drug. Finally, we used silver and antibiotic combinations in vitro to eradicate bacterial persister cells, and show both in vitro and in a mouse biofilm infection model that silver can enhance antibacterial action against bacteria that produce biofilms. This work shows that silver can be used to enhance the action of existing antibiotics against Gram-negative bacteria, thus strengthening the antibiotic arsenal for fighting bacterial infections.
The earth's limited resources demands our society their optimization so that the population's demands for energy, food, water, and medicines are fulfilled. Moreover, the expanding population, along ...with the socio-demographic shifts, and the emergence of new diseases, comprise the key challenges of this century. It is well-known that worldwide food and energy demands will duplicate by 2050, a scenario that would lead to the overexploitation of existing traditional animal and plant-based resources and cause adverse environmental impacts. Therefore, it is critical that new strategies should be pilared on sustainable production from alternative sustainable and environmentally friendly sources, to meet our society's long-term needs. Within the past decades, microalgae, as unicellular photosynthetic organisms, have attracted significant attention in various fields, from biofuel production to medical fields. Microalgae exhibit very interesting characteristics, such as their faster growth rate and higher biomass efficiency compared to plants. In addition, microalgae contain valuable bioresources, including lipids, pigments, carbohydrates, proteins, and bioactive compounds such as antioxidant, antibacterial, antiviral, anticancer, regenerative, antihypertensive, neuroprotective, and immunostimulating. However, the level of these compounds can be varied under different cultivation conditions such as nutrient availability, light intensity, CO2 concentration, temperature, and pH. There are various interesting reviews that have focused on different strategies to improve the growth and valuable compounds of microalgae, such as genetic engineering tools. However, recently, nanotechnology emerges as a field that offers novel and efficient methods to improve culturing conditions. To enhance the valuable compounds in microalgae, nanoparticles (NPs) with unique and different physiochemical behaviors are utilized and this review focuses on discussing and providing the state-of-the-art techniques that involve NPs and that are employed to improve microalgae growth rate, light utilization, access to carbon dioxide, and therefore increase the valuable compounds in microalgae cells.
•Nanoparticles have applications in gene expression regulation in microalgae.•Nanotechnology allows bioprocess production in microalgae.•Carbon dioxide fixation in microalgae enhanced using nanoparticles.
The world is facing a significant increase in infections caused by drug-resistant infectious agents. In response, various strategies have been recently explored to treat them, including the ...development of bacteriocins. Bacteriocins are a group of antimicrobial peptides produced by bacteria, capable of controlling clinically relevant susceptible and drug-resistant bacteria. Bacteriocins have been studied to be able to modify and improve their physicochemical properties, pharmacological effects, and biosafety. This manuscript focuses on the research being developed on the biosafety of bacteriocins, which is a topic that has not been addressed extensively in previous reviews. This work discusses the studies that have tested the effect of bacteriocins against pathogens and assess their toxicity using
models, including murine and other alternative animal models. Thus, this work concludes the urgency to increase and advance the
models that both assess the efficacy of bacteriocins as antimicrobial agents and evaluate possible toxicity and side effects, which are key factors to determine their success as potential therapeutic agents in the fight against infections caused by multidrug-resistant microorganisms.
Currently, the large amount of organic waste produced represents one of the leading environmental problems. According to United Nations, every year a third of all food produced ends up rotting and, ...therefore, becoming waste. Its use and revalorization represent an attractive alternative in the search for solutions to this situation. In this context, these residues can be used to obtain extracts beneficial for nanotechnology, acting as a reducing and stabilizing agent to produce metal nanomaterials such as silver nanoparticles by green synthesis, an ecofriendly and nontoxic method. Silver nanoparticles have been applied in applications as antibacterial, antifungal, and catalytic agents, as well as antioxidants, biosensors, and treatment of various diseases. Thus, this review contributes to fostering and promoting a sustainable environmental awareness through the reuse and revalorization of organic waste, which allows the sustainable biosynthesis of functional silver nanoparticles to be widely applied successfully in various fields of application.
The revalorization and use of organic waste allows obtaining extracts that act as a reducing and stabilizing agent for green and sustainable synthesis of functional AgNPs from Ag+ ions, which have potential application in biomedical and environmental areas due to their diverse properties such as antibacterial, antifungal, anti‐aging and treatment of various diseases, as well as catalytic agents, antioxidants, biosensors, among others.
With the increase in clinical cases of bacterial infections with multiple antibiotic resistance, the world has entered a health crisis. Overuse, inappropriate prescribing, and lack of innovation of ...antibiotics have contributed to the surge of microorganisms that can overcome traditional antimicrobial treatments. In 2017, the World Health Organization published a list of pathogenic bacteria, including
,
and
(ESKAPE). These bacteria can adapt to multiple antibiotics and transfer their resistance to other organisms; therefore, studies to find new therapeutic strategies are needed. One of these strategies is synthetic biology geared toward developing new antimicrobial therapies. Synthetic biology is founded on a solid and well-established theoretical framework that provides tools for conceptualizing, designing, and constructing synthetic biological systems. Recent developments in synthetic biology provide tools for engineering synthetic control systems in microbial cells. Applying protein engineering, DNA synthesis, and
design allows building metabolic pathways and biological circuits to control cellular behavior. Thus, synthetic biology advances have permitted the construction of communication systems between microorganisms where exogenous molecules can control specific population behaviors, induce intracellular signaling, and establish co-dependent networks of microorganisms.
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