Browsing by Author "Vargas Torres, Valentina Isabel"

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    Effect of recombinant protein production and release on microalgal fitness and the impact of environmental conditions for localized therapeutic delivery
    (Springer Nature, 2025) Carvajal Díaz, Felipe Alonso; Vargas Torres, Valentina Isabel; Becerra, Daniela; González Quezada, Nicolás Marcelo Orlando; Egaña, José T.
    Backgroud Genetically engineered photosynthetic microorganisms have been proposed as a therapeutic approach for the localized delivery of oxygen and recombinant proteins to tissues in various pathological conditions. However, the effect of recombinant protein production and secretion on microalgal fitness, as well as the impact of key environmental conditions on their potential therapeutic performance, has not yet been described. Therefore, in this study, the microalga Chlamydomonas reinhardtii was genetically engineered to produce and release the reporter protein mVenus and was then challenged by exposure to different media, temperatures, and substrates. Results The genetically modified microalgae were able to produce and release the mVenus protein under standard culture conditions without affecting overall fitness, including cell size and shape, growth potential, and oxygen metabolism, compared to the wild-type strain. Under mammalian cell culture conditions, the strains continued to produce and secrete mVenus protein for up to four days at 22 °C, 30 °C, and 37 °C. Additionally, photosynthetic biomaterials containing the engineered microalgae showed continuous recombinant protein release at 30 °C and 37 °C for up to four days. Conclusion The microalga Chlamydomonas reinhardtii can be genetically engineered to produce and release recombinant proteins without detrimental effects on its fitness, showing therapeutic potential under mammalian culture conditions and within biomaterials designed to promote tissue regeneration. Overall, these findings support the use of genetically engineered photosynthetic microalgae for the localized and controlled release of oxygen and recombinant proteins for several therapeutic applications.
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    Photosynthetic Solutions for Organ Perfusion Based on Microalgae and Cyanobacteria Display Differential In Vitro and In Vivo Features for Intravascular Oxygenation
    (American Chemical Society, 2025) Becerra, Daniela; Vargas Torres, Valentina Isabel; Veloso Giménez, Valentina del Carmen; Gallardo Agüero, Daniela; Miranda Cárdenas, Miguel Angel; Hernández Pavez, Valentina; Gonzalez Quezada, Nicolás Marcelo Orlando; San Martín, Sebastián; Boric P., Mauricio; Egaña, José T.
    The delivery of photosynthetic microorganisms has emerged as a strategy for tissue oxygenation, offering a promising approach to treat several hypoxic conditions. Among these, intravascular photosynthesis has been proposed for ex vivo organ preservation; however, the most suitable photosynthetic microorganisms and their behavior during intravascular perfusion remain to be fully elucidated. Therefore, this study evaluates key properties of photosynthetic solutions for organ perfusion, based on the microalgaeChlamydomonas reinhardtii and the cyanobacterium Synechococcus elongatus. In vitro characterization showed that both microorganisms maintained viability, morphology, and oxygen production capacity in a Ringer’s lactate-based medium for at least 24 h, with both photosynthetic solutions exhibiting rheological properties compatible with organ perfusion. In vivo perfusion of rat kidneys demonstrates sustained hemodynamic stability, with S. elongatus showing lower variability in vascular resistance. Histological analysis revealed significant retention of both microorganisms within renal structures, with S. elongatus inducing less tubular damage. Additionally, biocompatibility assays with human endothelial cells and zebrafish larvae showed no significant cytotoxic effects of the photosynthetic solutions. These findings support the feasibility of using photosynthetic microorganisms for intravascular photosynthesis, highlighting S. elongatus as particularly promising due to its lower oxygen consumption in darkness and reduced tissue damage after perfusion. This work provides significant insights toward the development of biologically active perfusion systems for innovative preservation strategies for organ transplantation.
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    Universal loop assembly: open, efficient and cross-kingdom DNA fabrication.
    (2020) Pollak, Bernardo; Matute Torres, Tamara Francisca; Núñez Quijada, Isaac Natán; Cerda Rojas, Ariel; López Sierra, Constanza Andrea; Vargas Torres, Valentina Isabel; Kan, Anton; Bielinski,Vincent; Dassow, Peter von; Dupont, Chris L.; Federici, Fernán
    Standardized type IIS DNA assembly methods are becoming essential for biological engineering and research. These methods are becoming widespread and more accessible due to the proposition of a 'common syntax' that enables higher interoperability between DNA libraries. Currently, Golden Gate (GG)-based assembly systems, originally implemented in hostspecific vectors, are being made compatible with multiple organisms. We have recently developed the GG-based Loop assembly system for plants, which uses a small library and an intuitive strategy for hierarchical fabrication of large DNA constructs (>30 kb). Here, we describe 'universal Loop' (uLoop) assembly, a system based on Loop assembly for use in potentially any organismof choice. This design permits the use of a compact number of plasmids (two sets of four odd and even vectors), which are utilized repeatedly in alternating steps. The elements required for transformation/maintenance in target organisms are also assembled as standardized parts, enabling customization of host-specific plasmids. Decoupling of the Loop assembly logic from the host-specific propagation elements enables universal DNA assembly that retains high efficiency regardless of the final host. As a proof-of-concept, we show the engineering of multigene expression vectors in diatoms, yeast, plants and bacteria. These resources are available through the OpenMTA for unrestricted sharing and open access.