As redesigning organisms using engineering principles is one of the purposes of synthetic biology (SynBio), the standardization of experimental methods and DNA parts is becoming increasingly a necessity. The synthetic biology community focusing on the engineering of Saccharomyces cerevisiae has been in the foreground in this area, conceiving several well-characterized SynBio toolkits widely adopted by the community. In this review, the molecular methods and toolkits developed for S. cerevisiae are discussed in terms of their contributions to the required standardization efforts. In addition, the toolkits designed for emerging nonconventional yeast species including Yarrowia lipolytica , Komagataella phaffii , and Kluyveromyces marxianus are also reviewed. Without a doubt, the characterized DNA parts combined with the standardized assembly strategies highlighted in these toolkits have greatly contributed to the rapid development of many metabolic engineering and diagnostics applications among others. Despite the growing capacity in deploying synthetic biology for common yeast genome engineering works, the yeast community has a long journey to go to exploit it in more sophisticated and delicate applications like bioautomation.

The alarming problem of meat adulteration emphasises the demand for accessible analytical approaches for food regulatory agencies to detect and, specially, to measure altered meat fractions. This study proposes a novel cross-species triplex droplet digital polymerase chain reaction (ddPCR) assay to simultaneously identify and quantify the ratios of pork/beef meat fractions from a total DNA content, including processed and autoclaved meat, without requiring a standard, achieving high sensitivity with a limit of quantification estimated at 0.1% (w/w) and a limit of detection down to 0.01% (w/w). A single copy nuclear gene, b-actin, was employed as a target, accompanied with myostatin gene as a cross-species target to quantify the meat background. The duplex assay provided a simultaneous quantification of pork and myostatin, whereas the triplex assay was able to detect pork, beef and myostatin with a decrease of technical error, cost and time.

Synthetic biology is a new area of science that operates at the intersection of engineering and biology and aims to design and synthesize living organisms and systems to perform new or improved functions. Despite the important role it plays in resolving global issues, instructing synthetic biology can be challenged by a limited availability of specific educational materials and techniques for explaining complex molecular mechanisms. On the other hand, digital fabrication tools, which allow the creation of 3D objects, are increasingly used for educational purposes, and several computational structures of molecular components commonly used in synthetic biology processes are deposited in open databases. Therefore, we hypothesized that the use of computer-assisted design (CAD) and 3D printing to create biomolecular structural models through hands-on interaction, followed by reflective observation, critical and analytical thinking, could enhance students’ learning in synthetic biology. In this sense, the present work describes the design, 3D printing process, and evaluation in classrooms of the molecular models of the first synthetic biological circuit, the genetic toggle switch. The 3D printed molecular structures can be freely downloaded and used by teachers to facilitate the training of STEM students in synthetic biology. Most importantly, the results demonstrated that our resource showed a significant positive impact (p < 0.05) on students’ learning process, indicating that the proposed method helped them better understand the genetic toggle switch.

Synthetic biology is a new area of science that operates at the intersection of engineering and biology and aims to design and synthesize living organisms and systems to perform new or improved functions. This novel biological area is considered of extreme relevance for the development of solutions to global problems. However, its teaching is often inaccessible to students, since many educational resources and methodological procedures are not available for understanding of complex molecular processes. On the other hand, digital fabrication tools, which allow the creation of 3D objects, are increasingly used for educational purposes, and several computational structures of molecular components commonly used in synthetic biology processes are deposited in open databases. Therefore, we hypothesize that the creation of biomolecular structures models by handling 3D physical objects using computer-assisted design (CAD) and additive fabrication based on 3D printing could help professors in synthetic biology teaching. In this sense, the present work describes the design and 3D print of the molecular models of the first synthetic genetic circuit, the toggle switch, which can be freely downloaded and used by teachers to facilitate the training of STEM students in synthetic biology.

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