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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    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.