New food sources and production systems:a comparison of international regulations and China’s advancements in novel foods with synthetic biology

The global shift towards sustainable food systems has sparked innovations in food sources and production systems,including cell-based meat,plant-based food products,precision fermentation,and 3D food printing.These advancements pose regulatory challenges and opportunities,with China emerging as a critical player in adopting and regulating new food technologies.This review explores the international landscape of new food sources and production systems(NFPS),focusing on China’s role and regulatory approaches compared to global practices.Through this comparative analysis,we aim to contribute to the ongoing dialogue on food safety regulation,offering insights and recommendations for policymakers,industry stakeholders,and researchers engaged in the global food system’s evolution.This comprehensive overview underscores the dynamic nature of regulatory frameworks governing NFPS,highlighting the international efforts to ensure food safety,consumer protection,and the sustainable evolution of the food industry.

Exploration on Teaching Reform of Synthetic Biology Course Based on the New Engineering Concept

Synthetic biology is a new frontier of life science,which aims to design,transform and even synthesize organisms with engineering design concept.Doing a good job in the teaching of"synthetic biology"is of great significance to the cultivation and reserve of biotechnology professionals in China,and also has an important impact on students' employment competitiveness.Under the background of"new engineering",the course reform of"synthetic biology"was carried out in terms of the construction of teaching staff,teaching methods,students' participation and the innovation of course content,and specific reform suggestions were put forward,hoping to effectively promote the sustainable development of"synthetic biology"and effectively improve the quality of education.

Food synthetic biology-driven protein supply transition: From animal-derived production to microbial fermentation

Animal-derived protein production is one of the major traditional protein supply methods,which continues to face increasing challenges to satisfy global needs due to population growth,augmented individual protein consumption,and aggravated environmental pollution.Thus,ensuring a sustainable protein source is a considerable challenge.The emergence and development of food synthetic biology has enabled the establishment of cell factories that effectively synthesize proteins,which is an important way to solve the protein supply problem.This review aims to discuss the existing problems of traditional protein supply and to elucidate the feasibility of synthetic biology in the process of protein synthesis.Moreover,using artificial bioengineered milk and artificial bioengineered eggs as examples,the progress of food protein supply transition based on synthetic biology has been systematically summarized.Additionally,the future of food synthetic biology as a potential source of protein has been also discussed.By strengthening and innovating the application of food synthetic biology technologies,including genetic engineering and high-throughput screening methods,the current limitations of artificial foods for protein synthesis and production should be addressed.Therefore,the development and industrial production of new food resources should be explored to ensure safe,high-quality,and sustainable global protein supply.

Efficient production of chemicals from microorganism by metabolic engineering and synthetic biology

The use of traditional chemical catalysis to produce chemicals has a series of drawbacks,such as high dependence on fossil resources,high energy consumption,and environmental pollution.With the development of synthetic biology and metabolic engineering,the use of renewable biomass raw materials for chemicals synthesis by constructing efficient microbial cell factories is a green way to replace traditional chemical catalysis and traditional microbial fermentation.This review mainly summarizes several types of bulk chemicals and high value-added chemicals using metabolic engineering and synthetic biology strategies to achieve efficient microbial production.In addition,this review also summarizes several strategies for effectively regulating microbial cell metabolism.These strategies can achieve the coupling balance of material and energy by regulating intracellular material metabolism or energy metabolism,and promote the efficient production of target chemicals by microorganisms.

Current advances of biocontainment strategy in synthetic biology

Synthetic biotechnology has led to the widespread application of genetically modified organisms(GMOs)in biochemistry, bioenergy, and therapy. However, the uncontrolled spread of GMOs may lead to genetic contamination by horizontal gene transfer, resulting in unpredictable biosafety risks. To deal with these challenges, many effective methods have been developed for biocontainment. In this article, we summarize and discuss recent advances in biocontainment strategies from three aspects: DNA replication, transcriptional regulation, and protein translation. We also briefly introduce the efforts in the biocontainment convention, such as the recent publication of the Tianjin Biosecurity Guidelines for the Code of Conduct for Scientists.

Cell-free synthetic biology in the new era of enzyme engineering

With the gradual rise of enzyme engineering,it has played an essential role in synthetic biology,medicine,and biomanufacturing.However,due to the limitation of the cell membrane,the complexity of cellular metabolism,the difficulty of controlling the reaction environment,and the toxicity of some metabolic products in traditional in vivo enzyme engineering,it is usually problematic to express functional enzymes and produce a high yield of synthesized compounds.Recently,cell-free synthetic biology methods for enzyme engineering have been proposed as alternative strategies.This cell-free method has no limitation of the cell membrane and no need to maintain cell viability,and each biosynthetic pathway is highly flexible.This property makes cell-free approaches suitable for the production of valuable products such as functional enzymes and chemicals that are difficult to synthesize.This article aims to discuss the latest advances in cell-free enzyme engineering,assess the trend of this developing topical filed,and analyze its prospects.

An international comprehensive benchmarking analysis of synthetic biology in China from 2015 to 2020

As a new interdisciplinary field,synthetic biology has led to valuable innovations in the fields of medicine,chemistry,agriculture,energy and environment.In this paper,we systematically review the development status of global synthetic biology in the past six years,and make an in-depth benchmarking analysis of the field in China.With the aid of Scopus and SciVal,we analyze the scholarly output of synthetic biology in the world and individual countries,including publication distribution,popular journals and eminent institutions.Furthermore,the research focus and concepts,citation impact and collaborations are also examined using numerical index methods such as the field-weighted citation impact(FWCI)and relative activity index(RAI),showing the differences between data more intuitively.This study aims to offer a comprehensive understanding of the research status of synthetic biology in China and the world,offering a benchmarked overview of the results as a reference to guide the development of this field in the future.

Synthetic Biology Construct of Ebola Virus in Bacteria Surrogate Is Stable and Safe for Rapid Detection Studies in a BSL-2 Laboratory Setting

Rapid detection of virulent pathogens during an outbreak is critical for public health advisories and control of the disease in a population. While many molecular techniques for point of care and clinical diagnosis abound, the US experience with the COVID-19 testing in the early stages of the pandemic underscores the critical importance of determining the appropriate target gene(s) with in-built controls that reliably detect pathogens with high sensitivity and specificity. Assays and research for diagnostics and therapy could be slowed during an epidemic because access to the required BSL-3 and BSL-4 laboratories are limited. So, during the 2014 West Africa Ebola outbreak, we tested the hypothesis that using synthetic cDNA of Ebolavirus in a bacteria surrogate (fit for all lab settings), would remain unmutated and safe after several generations, serving as an effective positive control in research settings, self test and point-of-care detection platforms. Primers were designed for the detection and quantification of the nucleoprotein (NP) gene of the 2014 Makona Ebola strain (KR781608.1, 733 - 1332 bp). To test the stability of artificially inserted translation arrest in the Orf of the model gene, it was edited to include three STOP codons in the RNA transcript using SNAP GENE. The segment was then spliced into a high copy number plasmid, cloned into One ShotTM TOP10 Escherichia coli (Invitrogen), and tested for stability and safety by periodic subculture, extraction and sequencing. Unlike COVID-19, rapid detection of blood-borne etiologies like Ebola requires optimized protocols for blood matrix. Using real-time PCR and newly designed primer pairs, the EBOV surrogate was detected and enumerated in human blood and regular broth and buffers. Based on aligned sequence analysis, the EBOV synthetic NP gene was stable (>99.9999% similarity coefficient) for at least 3 months. Detection sensitivity in broth and blood was at least 100 cells/ml or about 5.8 × 103 to 7.3 × 103 virion equivalents per ml. While the developments of transcription-and-replication-competent virus like particles (trVLP) have made it possible to study the infection and replication cycles of virulent pathogens in BSL-2 laboratories, the simplicity of our model and the reproducibility of detection and enumeration show the utility of synthetic bio-components as positive controls for point of care diagnostic tools. The inserted stop codons remained intact after many generations, suggesting that expressed virulent proteins can be easily silenced in synthetic biology models for research in BSL-1 and 2 and a wide range of pathogens. Synthetic bio-components can thereby aid further research by reducing costs and improving safety for workers and stakeholders.

Unique biogenesis and kinetics of hornwort Rubiscos revealed by synthetic biology systems

Hornworts are the only land plants that employ a pyrenoid to optimize Rubisco’s CO_(2) fixation,yet hornwort Rubisco remains poorly characterized.Here we assembled the hornwort Anthoceros agrestis Rubisco(AaRubisco)using the Arabidopsis thaliana SynBio expression system and observed the formation of stalled intermediates,prompting us to develop a new SynBio system with A.agrestis cognate chaperones.We successfully assembled AaRubisco and Rubisco from three other hornwort species.Unlike A.thaliana Rubisco,AaRubisco assembly is not dependent on RbcX or Raf2.Kinetic characterization reveals that hornwort Rubiscos exhibit a range of catalytic rates(3–10 s−1),but with similar affinity(∼30μM)and specificity(∼70)for CO_(2).These results suggest that hornwort Rubiscos do not comply with the long-held canonical catalytic trade-off observed in other land plants,providing experimental support that Rubisco kinetics may be phylogenetically constrained.Unexpectedly,we observed a 50%increase in AaRubisco catalytic rates when RbcX was removed from our SynBio system,without any reduction in specificity.Structural biology,biochemistry,and proteomic analysis suggest that subtle differences in Rubisco large-subunit interactions,when RbcX is absent during biogenesis,increases the accessibility of active sites and catalytic turnover rate.Collectively,this study uncovered a previously unknown Rubisco kinetic parameter space and provides a SynBio chassis to expand the survey of other Rubisco kinetics.Our discoveries will contribute to developing new approaches for engineering Rubisco with superior kinetics.

Synthetic biology:A powerful booster for future agriculture

Future agricultural development needs to solve the food security crisis caused by the global food shortage and the environmental demand for green and sustainable technology.The vigorous development of synthetic biology has brought new opportunities for modern agriculture.Synthetic biology can transform crops'metabolic pathways and genetic information and involves microorganisms'application in agriculture.Therefore,it has bright prospects in crop breeding and yield increase and ensuring the safety of the agricultural production environment.This perspective summarizes the application status and future development of synthetic biology in agriculture from the aspects of plant breeding,photosynthetic system,nitrogen fixation,and microorganisms.

Synthetic biology approaches for improving the specificity and efficacy of cancer immunotherapy

Immunotherapy has shown robust efficacy in treating a broad spectrum of hematological and solid cancers.Despite the transformative impact of immunotherapy on cancer treatment,several outstanding challenges remain.These challenges include on-target off-tumor toxicity,systemic toxicity,and the complexity of achieving potent and sustainable therapeutic efficacy.Synthetic biology has emerged as a promising approach to overcome these obstacles,offering innovative tools for engineering living cells with customized functions.This review provides an overview of the current landscape and future prospects of cancer immunotherapy,particularly emphasizing the role of synthetic biology in augmenting its specificity,controllability,and efficacy.We delineate and discuss two principal synthetic biology strategies:those targeting tumor surface antigens with engineered immune cells and those detecting intratumoral disease signatures with engineered gene circuits.This review concludes with a forwardlooking perspective on the enduring challenges in cancer immunotherapy and the potential breakthroughs that synthetic biology may contribute to the field.

Engineering the next generation of theranostic biomaterials with synthetic biology

Biomaterials have evolved from inert materials to responsive entities,playing a crucial role in disease diagnosis,treatment,and modeling.However,their advancement is hindered by limitations in chemical and mechanical approaches.Synthetic biology enabling the genetically reprograming of biological systems offers a new paradigm.It has achieved remarkable progresses in cell reprogramming,engineering designer cells for diverse applications.Synthetic biology also encompasses cell-free systems and rational design of biological molecules.This review focuses on the application of synthetic biology in theranostics,which boost rapid development of advanced biomaterials.We introduce key fundamental concepts of synthetic biology and highlight frontier applications thereof,aiming to explore the intersection of synthetic biology and biomaterials.This integration holds tremendous promise for advancing biomaterial engineering with programable complex functions.

An integrative database and its application for plant synthetic biology research

Plant synthetic biology research requires diverse bioparts that facilitate the redesign and construction of new-to-nature biological devices or systems in plants.Limited by few well-characterized bioparts for plant chassis,the development of plant synthetic biology lags behind that of its microbial counterpart.Here,we constructed a web-based Plant Synthetic BioDatabase(PSBD),which currently categorizes 1677 catalytic bioparts and 384 regulatory elements and provides information on 309 species and 850 chemicals.Online bioinformatics tools including local BLAST,chem similarity,phylogenetic analysis,and visual strength are provided to assist with the rational design of genetic circuits for manipulation of gene expression in planta.We demonstrated the utility of the PSBD by functionally characterizing taxadiene synthase 2 and its quan-titative regulation in tobacco leaves.More powerful synthetic devices were then assembled to amplify the transcriptional signals,enabling enhanced expression offlavivirus non-structure 1 proteins in plants.The PSBD is expected to be an integrative and user-centered platform that provides a one-stop service for diverse applications in plant synthetic biology research.

Regulatory RNAs in Bacillus subtilis:A review on regulatory mechanism and applications in synthetic biology

Bacteria exhibit a rich repertoire of RNA molecules that intricately regulate gene expression at multiple hierarchical levels,including small RNAs(sRNAs),riboswitches,and antisense RNAs.Notably,the majority of these regulatory RNAs lack or have limited protein-coding capacity but play pivotal roles in orchestrating gene expression by modulating transcription,post-transcription or translation processes.Leveraging and redesigning these regulatory RNA elements have emerged as pivotal strategies in the domains of metabolic engineering and synthetic biology.While previous investigations predominantly focused on delineating the roles of regulatory RNA in Gram-negative bacterial models such as Escherichia coli and Salmonella enterica,this review aims to summarize the mechanisms and functionalities of endogenous regulatory RNAs inherent to typical Gram-positive bacteria,notably Bacillus subtilis.Furthermore,we explore the engineering and practical applications of these regulatory RNA elements in the arena of synthetic biology,employing B.subtilis as a foundational chassis.

Engineering pluripotent stem cells with synthetic biology for regenerative medicine

Pluripotent stem cells(PSCs),characterized by self-renewal and capacity of differentiating into three germ layers,are the programmable building blocks of life.PSC-derived cells and multicellular systems,particularly organoids,exhibit great potential for regenerative medicine.However,this field is still in its infancy,partly due to limited strategies to robustly and precisely control stem cell behaviors,which are tightly regulated by inner gene regulatory networks in response to stimuli from the extracellular environment.Synthetic receptors and genetic circuits are powerful tools to customize the cellular sense-and-response process,suggesting their underlying roles in precise control of cell fate decision and function reconstruction.Herein,we review the progress and challenges needed to be overcome in the fields of PSC-based cell therapy and multicellular system generation,respectively.Furthermore,we summarize several well-established synthetic biology tools and their applications in PSC engineering.Finally,we highlight the challenges and perspectives of harnessing synthetic biology to PSC engineering for regenerative medicine.

Enabling technology and core theory of synthetic biology

Synthetic biology provides a new paradigm for life science research(“build to learn”)and opens the future journey of biotechnology(“build to use”).Here,we discuss advances of various principles and technologies in the mainstream of the enabling technology of synthetic biology,including synthesis and assembly of a genome,DNA storage,gene editing,molecular evolution and de novo design of function proteins,cell and gene circuit engineering,cell-free synthetic biology,artificial intelligence(AI)-aided synthetic biology,as well as biofoundries.We also introduce the concept of quantitative synthetic biology,which is guiding synthetic biology towards increased accuracy and predictability or the real rational design.We conclude that synthetic biology will establish its disciplinary system with the iterative development of enabling technologies and the maturity of the core theory.

Synthetic biology in the UK-An outline of plans and progress

Synthetic biology is capable of delivering new solutions to key challenges spanning the bioeconomy,both nationally and internationally.Recognising this significant potential and the associated need to facilitate its translation and commercialisation the UK government commissioned the production of a national Synthetic Biology Roadmap in 2011,and subsequently provided crucial support to assist its implementation.Critical infrastructural investments have been made,and important strides made towards the development of an effectively connected community of practitioners and interest groups.A number of Synthetic Biology Research Centres,DNA Synthesis Foundries,a Centre for Doctoral Training,and an Innovation Knowledge Centre have been established,creating a nationally distributed and integrated network of complementary facilities and expertise.The UK Synthetic Biology Leadership Council published a UK Synthetic Biology Strategic Plan in 2016,increasing focus on the processes of translation and commercialisation.Over 50 start-ups,SMEs and larger companies are actively engaged in synthetic biology in the UK,and inward investments are starting to flow.Together these initiatives provide an important foundation for stimulating innovation,actively contributing to international research and development partnerships,and helping deliver useful benefits from synthetic biology in response to local and global needs and challenges.

Synthetic biology identifies the minimal gene set required for paclitaxel biosynthesis in a plant chassis

The diterpenoid paclitaxel(Taxol)is a chemotherapy medication widely used as a first-line treatment against several types of solid cancers.The supply of paclitaxel from natural sources is limited.However,missing knowledge about the genes involved in several specific metabolic steps of paclitaxel biosynthesis has rendered it difficult to engineer the full pathway.In this study,we used a combination of transcriptomics,cell biology,metabolomics,and pathway reconstitution to identify the complete gene set required for the heterologous production of paclitaxel.We identified the missing steps from the current model of paclitaxel biosynthesis and confirmed the activity of most of the missing enzymes via heterologous expression in Nicotiana benthamiana.Notably,we identified a new C4β-C20 epoxidase that could overcome the first bottleneck of metabolic engineering.We used both previously characterized and newly identified oxomutases/epoxidases,taxane 1β-hydroxylase,taxane 9aα-hydroxylase,taxane 9α-dioxygenase,and phenylalanine-CoA ligase,to successfully biosynthesize the key intermediate baccatin Ill and to convert baccatin Ill into paclitaxel in N.benthamiana.In combination,these approaches establisha metabolic route to taxoidbiosynthesis and provide insights into the unique chemistry that plants use to generate complex bioactive metabolites.

Synthetic biology for CO2 fixation

Recycling of carbon dioxide(CO_2) into fuels and chemicals is a potential approach to reduce CO_2 emission and fossil-fuel consumption. Autotrophic microbes can utilize energy from light, hydrogen, or sulfur to assimilate atmospheric CO_2 into organic compounds at ambient temperature and pressure. This provides a feasible way for biological production of fuels and chemicals from CO_2 under normal conditions. Recently great progress has been made in this research area, and dozens of CO_2-derived fuels and chemicals have been reported to be synthesized by autotrophic microbes. This is accompanied by investigations into natural CO_2-fixation pathways and the rapid development of new technologies in synthetic biology. This review first summarizes the six natural CO_2-fixation pathways reported to date, followed by an overview of recent progress in the design and engineering of CO_2-fixation pathways as well as energy supply patterns using the concept and tools of synthetic biology. Finally, we will discuss future prospects in biological fixation of CO_2.

Development of synthetic biology tools to engineer Pichia pastoris as a chassis for the production of natural products

The methylotrophic yeast Pichia pastoris(a.k.a.Komagataella phaffii)is one of the most commonly used hosts for industrial production of recombinant proteins.As a non-conventional yeast,P.pastoris has unique biological characteristics and its expression system has been well developed.With the advances in synthetic biology,more efforts have been devoted to developing P.pastoris into a chassis for the production of various high-value compounds,such as natural products.This review begins with the introduction of synthetic biology tools for the engineering of P.pastoris,including vectors,promoters,and terminators for heterologous gene expression as well as Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated System(CRISPR/Cas)for genome editing.This review is then followed by examples of the production of value-added natural products in metabolically engineered P.pastoris strains.Finally,challenges and outlooks in developing P.pastoris as a synthetic biology chassis are prospected.