Synthetic biology (Oxford, England)最新文献

{"title":"Moving towards chemical-free agriculture, 37 kb at a time.","authors":"Sonja Billerbeck","doi":"10.1093/synbio/ysab009","DOIUrl":"https://doi.org/10.1093/synbio/ysab009","url":null,"abstract":"Domestic crop plants are modern marvels of extensive breeding; however, many of their natural defenses against pests and pathogens have been lost. Wild relatives still harbor disease resistance genes, but transferring these large sequences into complex, polyploid plant genomes calls for advanced genomic engineering technologies. Recently, government researchers in Australia, successfully transferred a 37kb resistance stack into the genome of a domesticated wheat species such that it is pro-tected against the rapidly evolving wheat leaf rust pathogen Puccinia graminis f. sp. tritici ( Pgt ) without losing any agronomic features. 1 Plant diseases caused by pathogenic fungi can devastate crop yield and pose a threat to food security. 2,3 About 30% of our most important crops are lost every year to fungal diseases. 3 Over decades, agricultural crops have been bred towards maxi-mum productivity under high fungicide treatment, meanwhile breeding out the plants’ own defense genes. 3 The genetic ar-mory still intact in wild crop relatives (so-called R genes) could provide an effective means towards a chemical-free disease control. 4 Introducing those genes into domestic crops is a multi-factorial challenge yet underappreciated by much of the synthetic biology community.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysab009"},"PeriodicalIF":0.0,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7937910/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25488216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptional control of <i>Clostridium autoethanogenum</i> using CRISPRi.","authors":"Nicholas Fackler,&nbsp;James Heffernan,&nbsp;Alex Juminaga,&nbsp;Damien Doser,&nbsp;Shilpa Nagaraju,&nbsp;R Axayacatl Gonzalez-Garcia,&nbsp;Séan D Simpson,&nbsp;Esteban Marcellin,&nbsp;Michael Köpke","doi":"10.1093/synbio/ysab008","DOIUrl":"https://doi.org/10.1093/synbio/ysab008","url":null,"abstract":"<p><p>Gas fermentation by <i>Clostridium autoethanogenum</i> is a commercial process for the sustainable biomanufacturing of fuels and valuable chemicals using abundant, low-cost C1 feedstocks (CO and CO₂) from sources such as inedible biomass, unsorted and nonrecyclable municipal solid waste, and industrial emissions. Efforts toward pathway engineering and elucidation of gene function in this microbe have been limited by a lack of genetic tools to control gene expression and arduous genome engineering methods. To increase the pace of progress, here we developed an inducible CRISPR interference (CRISPRi) system for <i>C. autoethanogenum</i> and applied that system toward transcriptional repression of genes with ostensibly crucial functions in metabolism.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysab008"},"PeriodicalIF":0.0,"publicationDate":"2021-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysab008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38853817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Joint universal modular plasmids (JUMP): a flexible vector platform for synthetic biology.","authors":"Marcos Valenzuela-Ortega,&nbsp;Christopher French","doi":"10.1093/synbio/ysab003","DOIUrl":"https://doi.org/10.1093/synbio/ysab003","url":null,"abstract":"<p><p>Generation of new DNA constructs is an essential process in modern life science and biotechnology. Modular cloning systems based on Golden Gate cloning, using Type IIS restriction endonucleases, allow assembly of complex multipart constructs from reusable basic DNA parts in a rapid, reliable and automation-friendly way. Many such toolkits are available, with varying degrees of compatibility, most of which are aimed at specific host organisms. Here, we present a vector design which allows simple vector modification by using modular cloning to assemble and add new functions in secondary sites flanking the main insertion site (used for conventional modular cloning). Assembly in all sites is compatible with the PhytoBricks standard, and vectors are compatible with the Standard European Vector Architecture (SEVA) as well as BioBricks. We demonstrate that this facilitates the construction of vectors with tailored functions and simplifies the workflow for generating libraries of constructs with common elements. We have made available a collection of vectors with 10 different microbial replication origins, varying in copy number and host range, and allowing chromosomal integration, as well as a selection of commonly used basic parts. This design expands the range of hosts which can be easily modified by modular cloning and acts as a toolkit which can be used to facilitate the generation of new toolkits with specific functions required for targeting further hosts.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysab003"},"PeriodicalIF":0.0,"publicationDate":"2021-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysab003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25404686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Starting from scratch: a workflow for building truly novel proteins.","authors":"Pablo Cárdenas","doi":"10.1093/synbio/ysab005","DOIUrl":"https://doi.org/10.1093/synbio/ysab005","url":null,"abstract":"In its early days, synthetic biology was often defined as repurposing existing biological parts for new applications. More recently, we have seen work that pushes the boundaries of the field past repurposing and into the design of truly novel biological parts. To date, most attempts at designing complex, non-structural protein functions have hinged on grafting protein motifs with known functions onto synthetic protein scaffolds. This ‘top-down’ approach to synthetic protein design unfortunately depends on the structural compatibility of the functional sites with rigid scaffolds used. A study recently published in Nature Chemical Biology describes an alternative ‘bottom-up’ approach in which structural elements are created de novo to support the functional elements in whatever conformation they are specified in (1). The multi-institutional team led by Bruno Correira’s lab at École Polytechnique Fédérale de Lausanne demonstrated the efficacy of their novel approach by building tunable biosensors for epitope-specific antibodies and dual-specific ligands for synthetic cell receptors. To achieve this ‘bottom-up’ design, the authors used TopoBuilder (2), a previously published computational tool, to generate three-dimensional ‘sketches’ of a given protein of interest with specific functional motifs in their idealized conformations. With the help of another software tool, Rosetta FunFoldDes (3), the authors created and simulated tens of thousands of possible designs fulfilling the design criteria, which were filtered by favorable thermodynamic predictions for stability and folding. A combinatorial library of up to 10 million variants was built from elements of the best designs. The libraries were then screened by binding affinity to the desired target(s) and protease digestion using yeast surface display, and the best protein variants were selected by sequencing the output. Finally, the structure and behavior of the final products were determined using a variety of different physical and chemical assays. The authors used their workflow to design a biosensor based on bioluminescence resonant energy transfer (4) to sense antibodies with affinities for a single, specific epitope found in respiratory syncytial virus and metapneumovirus, two respiratory pathogens. The novel design pipeline made it possible to present the single epitope in scaffolds with different binding affinities to the target antibody. This, in turn, allowed for tuning the biosensor’s response. Furthermore, the workflow was used to create a synthetic ligand capable of binding to two different, previously created synthetic mammalian signal receptors, which trigger expression of a reporter gene (5). The authors demonstrated the ligand simultaneously bound its two distinct targets by showing it only triggered output signal when both types of receptor were present in a cell. The work presented by Yang et al. is exciting for its practically universal applicability in biological research. Appli","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysab005"},"PeriodicalIF":0.0,"publicationDate":"2021-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7966779/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25510053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aquarium: open-source laboratory software for design, execution and data management.","authors":"Justin Vrana,&nbsp;Orlando de Lange,&nbsp;Yaoyu Yang,&nbsp;Garrett Newman,&nbsp;Ayesha Saleem,&nbsp;Abraham Miller,&nbsp;Cameron Cordray,&nbsp;Samer Halabiya,&nbsp;Michelle Parks,&nbsp;Eriberto Lopez,&nbsp;Sarah Goldberg,&nbsp;Benjamin Keller,&nbsp;Devin Strickland,&nbsp;Eric Klavins","doi":"10.1093/synbio/ysab006","DOIUrl":"https://doi.org/10.1093/synbio/ysab006","url":null,"abstract":"<p><p>Automation has been shown to improve the replicability and scalability of biomedical and bioindustrial research. Although the work performed in many labs is repetitive and can be standardized, few academic labs can afford the time and money required to automate their workflows with robotics. We propose that human-in-the-loop automation can fill this critical gap. To this end, we present Aquarium, an open-source, web-based software application that integrates experimental design, inventory management, protocol execution and data capture. We provide a high-level view of how researchers can install Aquarium and use it in their own labs. We discuss the impacts of the Aquarium on working practices, use in biofoundries and opportunities it affords for collaboration and education in life science laboratory research and manufacture.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysab006"},"PeriodicalIF":0.0,"publicationDate":"2021-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysab006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39250784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing synthetic biology to expand chemical diversity of antibiotics.","authors":"Fatima Enam","doi":"10.1093/synbio/ysaa029","DOIUrl":"https://doi.org/10.1093/synbio/ysaa029","url":null,"abstract":"Antibiotic resistance is one of the greatest challenges facing public health. Historically, glycopeptide antibiotics (GPA) have been used in the treatment of infections caused by Grampositive pathogens, including multiresistant Staphylococcus aureus (MRSA) infections and enterococcal infections, which are resistant to beta-lactams and other antibiotics. Rising GPA resistance has led to the discovery and clinical development of synthetic second-generation glycopeptides but more needs to be done to stand up against the evolution of resistance mechanisms. In a recent publication in Nature Communications, a team of scientists led by Gerard Wright at McMaster University in Canada, developed a clever synthetic biology platform for the production and discovery of novel GPAs. GPAs are heptapeptides that are naturally synthesized in actinobacteria, orchestrated by co-localized genes in the genome, called biosynthetic gene clusters (BGCs), and a host of other genes for modification, regulation and transport. Although next-generation sequencing technologies and machine-learning tools have enabled the identification of a wealth of BGCs in bacterial genomes, low or no yields of GPAs from parental actinomycetes, lack of heterologous hosts for expressing BGCs and difficulty in cloning these large constructs (&gt;70 kb) has kept these GPAs elusive. Xu et al. engineered Streptomyces coelicolor, a genetically tractable microorganism, that contains the biosynthetic machinery and precursors required for GPA biosynthesis, including pathways for nonproteinogenic amino acid components to create the GPAHex chassis. To clone the large BGCs and increase selection efficiency, they developed an optimized transformation associated recombination (TAR) system that allows isolation and manipulation of large DNA constructs. The TAR system relies on a copy number control replicon that can be conditionally induced for high copy numbers. The system utilizes a ura3 counter-selection marker, which was also optimized by introducing additional transcriptional initiation sites between the TATA box and the start codon to maximize ura3 transcription. To demonstrate the synthesis of a GPA, they targeted corbomycin, a Type V GPA. Corbomycin was discovered by phylogeny guided genome mining, but further development was hindered by low titers in the producer Streptomyces strain. Xu et al. cloned a 76 kb region, constituting the peptide scaffold and six TISs, into the S. coelicolor chassis strain via E. coli-yeast triparental mating. Growth inhibition was observed against Bacillus subtilis and the production titer was 65.4 mg/l, 19-fold higher compared to the parental Streptomyces strain. The authors also used the GPAHex platform for the discovery of a GPA in an Amycolatopsis strain that shares homology with the teicoplanin class of GPAs but is transcriptionally inactive in the parental strain. They cloned the scaffold including four TISs into the chassis strain to express GP1416 that displayed an","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysaa029"},"PeriodicalIF":0.0,"publicationDate":"2021-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8056980/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38853816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis of libraries and multi-site mutagenesis using a PCR-derived, dU-containing template.","authors":"Gretchen Meinke,&nbsp;Nahide Dalda,&nbsp;Benjamin S Brigham,&nbsp;Andrew Bohm","doi":"10.1093/synbio/ysaa030","DOIUrl":"https://doi.org/10.1093/synbio/ysaa030","url":null,"abstract":"<p><p>Directed DNA libraries are useful because they focus genetic diversity in the most important regions within a sequence. Ideally, all sequences in such libraries should appear with the same frequency and there should be no significant background from the starting sequence. These properties maximize the number of different sequences that can be screened. Described herein is a method termed SLUPT (Synthesis of Libraries via a dU-containing PCR-derived Template) for generating highly targeted DNA libraries and/or multi-site mutations wherein the altered bases may be widely distributed within a target sequence. This method is highly efficient and modular. Moreover, multiple distinct sites, each with one or more base changes, can be altered in a single reaction. There is very low background from the starting sequence, and SLUPT libraries have similar representation of each base at the positions selected for variation. The SLUPT method utilizes a single-stranded dU-containing DNA template that is made by polymerase chain reaction (PCR). Synthesis of the template in this way is significantly easier than has been described earlier. A series of oligonucleotide primers that are homologous to the template and encode the desired genetic diversity are extended and ligated in a single reaction to form the mutated product sequence or library. After selective inactivation of the template, only the product library is amplified. There are no restrictions on the spacing of the mutagenic primers except that they cannot overlap.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"6 1","pages":"ysaa030"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8260824/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9770036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Building a biofoundry.","authors":"Maciej B Holowko,&nbsp;Emma K Frow,&nbsp;Janet C Reid,&nbsp;Michelle Rourke,&nbsp;Claudia E Vickers","doi":"10.1093/synbio/ysaa026","DOIUrl":"https://doi.org/10.1093/synbio/ysaa026","url":null,"abstract":"<p><p>A biofoundry provides automation and analytics infrastructure to support the engineering of biological systems. It allows scientists to perform synthetic biology and aligned experimentation on a high-throughput scale, massively increasing the solution space that can be examined for any given problem or question. However, establishing a biofoundry is a challenging undertaking, with numerous technical and operational considerations that must be addressed. Using collated learnings, here we outline several considerations that should be addressed prior to and during establishment. These include drivers for establishment, institutional models, funding and revenue models, personnel, hardware and software, data management, interoperability, client engagement and biosecurity issues. The high cost of establishment and operation means that developing a long-term business model for biofoundry sustainability in the context of funding frameworks, actual and potential client base, and costing structure is critical. Moreover, since biofoundries are leading a conceptual shift in experimental design for bioengineering, sustained outreach and engagement with the research community are needed to grow the client base. Recognition of the significant, long-term financial investment required and an understanding of the complexities of operationalization is critical for a sustainable biofoundry venture. To ensure state-of-the-art technology is integrated into planning, extensive engagement with existing facilities and community groups, such as the Global Biofoundries Alliance, is recommended.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":" ","pages":"ysaa026"},"PeriodicalIF":0.0,"publicationDate":"2020-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa026","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25559593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single enzyme RT-PCR of full-length ribosomal RNA.","authors":"Michael J Hammerling, Danielle J Yoesep, Michael C Jewett","doi":"10.1093/synbio/ysaa028","DOIUrl":"10.1093/synbio/ysaa028","url":null,"abstract":"<p><p>The ribosome is a two-subunit, macromolecular machine composed of RNA and proteins that carries out the polymerization of α-amino acids into polypeptides. Efforts to engineer ribosomal RNA (rRNA) deepen our understanding of molecular translation and provide opportunities to expand the chemistry of life by creating ribosomes with altered properties. Toward these efforts, reverse transcription PCR (RT-PCR) of the entire 16S and 23S rRNAs, which make up the 30S small subunit and 50S large subunit, respectively, is important for isolating desired phenotypes. However, reverse transcription of rRNA is challenging due to extensive secondary structure and post-transcriptional modifications. One key challenge is that existing commercial kits for RT-PCR rely on reverse transcriptases that lack the extreme thermostability and processivity found in many commercial DNA polymerases, which can result in subpar performance on challenging templates. Here, we develop methods employing a synthetic thermostable reverse transcriptase (RTX) to enable and optimize RT-PCR of the complete <i>Escherichia coli</i> 16S and 23S rRNAs. We also characterize the error rate of RTX when traversing the various post-transcriptional modifications of the 23S rRNA. We anticipate that this work will facilitate efforts to study and characterize many naturally occurring long RNAs and to engineer the translation apparatus for synthetic biology.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa028"},"PeriodicalIF":0.0,"publicationDate":"2020-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/05/df/ysaa028.PMC7772474.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39138445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthetic promoters went green: MinSyns bridge the gap between tunable expression and synthetic biology in plants.","authors":"Andrea Tagliani","doi":"10.1093/synbio/ysaa027","DOIUrl":"https://doi.org/10.1093/synbio/ysaa027","url":null,"abstract":"Precise control of gene expression is critical to allow the design of tunable synthetic gene circuits. To date, our ability to precisely predict orthogonal expression in plants lags well behind that of animals and bacteria. This is largely because traditional attempts to characterize plant promoters have found few reliable sequence patterns. Even the TATA box is found in a minority of plant promoters (1). As a result, plant biologists are still relying on a set of promoters that are not completely orthogonal and that often cannot ensure homogenous expression between different tissues; moreover, the length of these promoters, the local DNA environment of the insertion and other unknown factors often do not allow for tunable and specific expression. Recently, where classical approaches aimed at characterizing a discrete set of cis elements in plant promoters have failed to provide a comprehensive answer, machine learning helped researchers to blaze a new path for synthetic promoter design. In a paper published in Nucleic Acid Research, Cai et al. (2) developed a set of minimal synthetic promoters (MinSyns) by mining from typical promoters used in plant biology a set of rules by which, through a computational approach, the researchers were able to build a set of small standardized cis-regulatory elements (CREs) exploitable for green synthetic biology. The main fodder for the author’s machine learning approach consisted not of plant promoters, but of sequences derived from pathogenic plant viruses, which are widely used in plant biology to drive constitutive expression. They found that small CREs from a set of these promoters are regulated by an endogenous plant transcription factor. Indeed, deletion experiments confirmed the importance of these CREs in regulating expression. The authors used the experimentally determined strength of these CREs to generate a quantitative score for each. They then developed a script, which randomly assembles minimal promoters composed of few CREs for which a score is assigned, based on the relative promoter strength. Using luciferase-based reporter assays, the authors first confirmed that their MinSyn library could be exploited to tune transient expression in plant protoplasts. The authors then selected four MinSyns for further characterization in transgenic plants. MinSyn promoters predictably drove the constitutive expression of GUS or YFP in stable transgenic lines of Arabidopsis thaliana, Brassica rapa and Nicotiana benthamiana plants. Finally, the authors demonstrated that it is possible to build synthetic genetic circuits from MinSyns, allowing tunable expression of two genes and variable expression patterns depending on the number of cognate binding sites for an orthogonal TF. The novelty of this work lies in the organisms themselves. Due to their capacity to produce secondary metabolites and photosynthetic abilities, plants are arguably the most suitable chassis for the production of drugs, sustainable foods, ","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa027"},"PeriodicalIF":0.0,"publicationDate":"2020-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa027","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39443998","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}