ACS Synthetic Biology最新文献

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Biological Switches: Past and Future Milestones of Transcription Factor-Based Biosensors.
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-22 DOI: 10.1021/acssynbio.4c00689
Brecht De Paepe, Marjan De Mey
{"title":"Biological Switches: Past and Future Milestones of Transcription Factor-Based Biosensors.","authors":"Brecht De Paepe, Marjan De Mey","doi":"10.1021/acssynbio.4c00689","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00689","url":null,"abstract":"<p><p>Since the description of the <i>lac</i> operon in 1961 by Jacob and Monod, transcriptional regulation in prokaryotes has been studied extensively and has led to the development of transcription factor-based biosensors. Due to the broad variety of detectable small molecules and their various applications across biotechnology, biosensor research and development have increased exponentially over the past decades. Throughout this period, key milestones in fundamental knowledge, synthetic biology, analytical tools, and computational learning have led to an immense expansion of the biosensor repertoire and its application portfolio. Over the years, biosensor engineering became a more multidisciplinary discipline, combining high-throughput analytical tools, DNA randomization strategies, forward engineering, and advanced protein engineering workflows. Despite these advances, many obstacles remain to fully unlock the potential of biosensor technology. This review analyzes the timeline of key milestones on fundamental research (1960s to 2000s) and engineering strategies (2000s onward), on both the DNA and protein level of biosensors. Moreover, insights into the future perspectives, remaining hurdles, and unexplored opportunities of this promising field are discussed.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A Novel Membrane-Associated Protein Aids Bacterial Colonization of Maize.
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-21 DOI: 10.1021/acssynbio.4c00489
Maya Venkataraman, Valentina Infante, Grzegorz Sabat, Kai Sanos-Giles, Jean-Michel Ané, Brian F Pfleger
{"title":"A Novel Membrane-Associated Protein Aids Bacterial Colonization of Maize.","authors":"Maya Venkataraman, Valentina Infante, Grzegorz Sabat, Kai Sanos-Giles, Jean-Michel Ané, Brian F Pfleger","doi":"10.1021/acssynbio.4c00489","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00489","url":null,"abstract":"<p><p>The soil environment affected by plant roots and their exudates, termed the rhizosphere, significantly impacts crop health and is an attractive target for engineering desirable agricultural traits. Engineering microbes in the rhizosphere is one approach to improving crop yields that directly minimizes the number of genetic modifications made to plants. Soil microbes have the potential to assist with nutrient acquisition, heat tolerance, and drought response if they can persist in the rhizosphere in the correct numbers. Unfortunately, the mechanisms by which microbes adhere and persist on plant roots are poorly understood, limiting their application. This study examined the membrane proteome shift upon adherence to roots in two bacteria of interest, <i>Klebsiella variicola</i> and <i>Pseudomonas putida.</i> From this surface proteome data, we identified a novel membrane protein from a nonlaboratory isolate of <i>P. putida</i> that increases binding to maize roots using unlabeled proteomics. When this protein was moved from the environmental isolate to a common lab strain (<i>P. putida</i> KT2440), we observed increased binding capabilities of <i>P. putida</i> KT2440 to both abiotic mimic surfaces and maize roots. We observed a similar increased binding capability to maize roots when the protein was heterologously expressed in <i>K. variicola</i> and <i>Stutzerimonas stutzeri</i>. With the discovery of this novel binding protein, we outline a strategy for harnessing natural selection and wild isolates to build more persistent strains of bacteria for field applications and plant growth promotion.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Sequencing Strategy to Ensure Accurate Plasmid Assembly. 确保质粒组装准确的测序策略
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-11-07 DOI: 10.1021/acssynbio.4c00539
Sarah I Hernandez, Casey-Tyler Berezin, Katie M Miller, Samuel J Peccoud, Jean Peccoud
{"title":"Sequencing Strategy to Ensure Accurate Plasmid Assembly.","authors":"Sarah I Hernandez, Casey-Tyler Berezin, Katie M Miller, Samuel J Peccoud, Jean Peccoud","doi":"10.1021/acssynbio.4c00539","DOIUrl":"10.1021/acssynbio.4c00539","url":null,"abstract":"<p><p>Despite the wide use of plasmids in research and clinical production, the need to verify plasmid sequences is a bottleneck that is too often underestimated in the manufacturing process. Although sequencing platforms continue to improve, the method and assembly pipeline chosen still influence the final plasmid assembly sequence. Furthermore, few dedicated tools exist for plasmid assembly, especially for <i>de novo</i> assembly. Here, we evaluated short-read, long-read, and hybrid (both short and long reads) <i>de novo</i> assembly pipelines across three replicates of a 24-plasmid library. Consistent with previous characterizations of each sequencing technology, short-read assemblies had issues resolving GC-rich regions, and long-read assemblies commonly had small insertions and deletions, especially in repetitive regions. The hybrid approach facilitated the most accurate, consistent assembly generation and identified mutations relative to the reference sequence. Although Sanger sequencing can be used to verify specific regions, some GC-rich and repetitive regions were difficult to resolve using any method, suggesting that easily sequenced genetic parts should be prioritized in the design of new genetic constructs.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"4099-4109"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
De Novo Production of the Bioactive Phenylpropanoid Artepillin C Using Membrane-Bound Prenyltransferase in Komagataella phaffii. 利用 Komagataella phaffii 中的膜结合异戊烯基转移酶从新生产具有生物活性的苯丙氨酸青蒿素 C。
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-11-12 DOI: 10.1021/acssynbio.4c00472
Takahiro Bamba, Ryosuke Munakata, Yuya Ushiro, Ryota Kumokita, Sayaka Tanaka, Yoshimi Hori, Akihiko Kondo, Kazufumi Yazaki, Tomohisa Hasunuma
{"title":"De Novo Production of the Bioactive Phenylpropanoid Artepillin C Using Membrane-Bound Prenyltransferase in <i>Komagataella phaffii</i>.","authors":"Takahiro Bamba, Ryosuke Munakata, Yuya Ushiro, Ryota Kumokita, Sayaka Tanaka, Yoshimi Hori, Akihiko Kondo, Kazufumi Yazaki, Tomohisa Hasunuma","doi":"10.1021/acssynbio.4c00472","DOIUrl":"10.1021/acssynbio.4c00472","url":null,"abstract":"<p><p>Artepillin C is a diprenylated phenylpropanoid with various pharmacological benefits for human health. Its natural occurrence is limited to a few Asteraceae plants, such as <i>Baccharis</i> species, necessitating a stable supply through synthetic biology. In <i>Saccharomyces cerevisiae</i>, the utilization of aromatic substrates within the cell was limited, resulting in very low production of artepillin C. In this study, we used AcPT1, a <i>p</i>-coumaric acid (<i>p</i>-CA)-specific diprenyltransferase, in <i>Komagataella phaffii</i> to produce artepillin C. Detailed studies revealed that the critical bottleneck in <i>K. phaffii</i> was the supply of prenyl diphosphates, not phenylpropanoid flux. By enhancing the prenyl substrate pathway through overexpression of isopentenyl diphosphate isomerase and a truncated HMG-CoA reductase, we achieved a strong increase in artepillin C production. A major part of artepillin C was accumulated in yeast cells. One of the advantages of <i>K. phaffii</i> is its superior growth and ability to achieve high cell density cultivation compared to that of <i>S. cerevisiae</i>. Therefore, fed-batch cultivation with glycerol was performed. As a result, the dry cell weight (DCW) reached 61.0 g/L, and the intracellular amount of de novo produced artepillin C reached 187.3 μg/DCW. Analysis of intermediates revealed that the supply of <i>p</i>-CA constituted a bottleneck in artepillin C production in the engineered strain. By enhancing the <i>p</i>-CA supply, the intracellular accumulation of artepillin C reached 1200 μg/DCW even in batch cultivation. Moreover, the total intra- and extracellular amounts of artepillin C reached 12.5 mg/L, marking the highest de novo synthesis amount of artepillin C reported thus far, even under batch cultivation conditions.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"4040-4049"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142612581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Diagnostic and Therapeutic Microbial Circuit with Application to Intestinal Inflammation. 应用于肠道炎症的诊断和治疗微生物回路。
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-11-28 DOI: 10.1021/acssynbio.3c00668
Liana N Merk, Andrey S Shur, Smrutiti Jena, Javier Munoz, Douglas K Brubaker, Richard M Murray, Leopold N Green
{"title":"Diagnostic and Therapeutic Microbial Circuit with Application to Intestinal Inflammation.","authors":"Liana N Merk, Andrey S Shur, Smrutiti Jena, Javier Munoz, Douglas K Brubaker, Richard M Murray, Leopold N Green","doi":"10.1021/acssynbio.3c00668","DOIUrl":"10.1021/acssynbio.3c00668","url":null,"abstract":"<p><p>Bacteria genetically engineered to execute defined therapeutic and diagnostic functions in physiological settings can be applied to colonize the human microbiome, providing in situ surveillance and conditional disease modulation. However, many engineered microbes can only respond to single-input environmental factors, limiting their tunability, precision, and effectiveness as living diagnostic and therapeutic systems. For engineering microbes to improve complex chronic disorders such as inflammatory bowel disease, the bacteria must respond to combinations of stimuli in the proper context and time. This work implements a previously characterized split activator AND logic gate in the probiotic <i>Escherichia coli</i> strain Nissle 1917 (EcN). Our system can respond to two input signals: the inflammatory biomarker tetrathionate and a second input signal, anhydrotetracycline (aTc), for manual control. We report 4-6 fold induction with a minimal leak when the two chemical signals are present. We model the AND gate dynamics using chemical reaction networks and tune parameters in silico to identify critical perturbations that affect our circuit's selectivity. Finally, we engineer the optimized AND gate to secrete a therapeutic anti-inflammatory cytokine IL-22 using the hemolysin secretion pathway in the probiotic <i>E. coli</i> strain. We used a germ-free transwell model of the human gut epithelium to show that our engineering bacteria produce similar host cytokine responses compared to recombinant cytokine. Our study presents a scalable workflow to engineer cytokine-secreting microbes driven by logical signal processing. It demonstrates the feasibility of IL-22 derived from probiotic EcN with minimal off-target effects in a gut epithelial context.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"3885-3896"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142737748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Antibiotics-Free Steady Bioproduction of Valuable Chemicals from Organic Wastes by Engineered Vibrio natriegens through Targeted Gene Integration.
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-12-03 DOI: 10.1021/acssynbio.4c00669
Dan Sun, Hui-Hui Li, Jing Wu, Jie Wu, Wei-Qiang Lin, Ru-Li He, Dong-Feng Liu, Wen-Wei Li
{"title":"Antibiotics-Free Steady Bioproduction of Valuable Chemicals from Organic Wastes by Engineered <i>Vibrio natriegens</i> through Targeted Gene Integration.","authors":"Dan Sun, Hui-Hui Li, Jing Wu, Jie Wu, Wei-Qiang Lin, Ru-Li He, Dong-Feng Liu, Wen-Wei Li","doi":"10.1021/acssynbio.4c00669","DOIUrl":"10.1021/acssynbio.4c00669","url":null,"abstract":"<p><p>Bioproduction of chemicals by using engineered bacteria is promising for a circular economy but challenged the instability of the introduced plasmid by conventional methods. Here, we developed a two-plasmid INTEGRET system to reliably integrate the targeted gene into the <i>Vibrio natriegens</i> genome, making it a powerful strain for efficient and steady bioproduction without requiring antibiotic addition. The INTEGRET system allows for gene insertion at over 75% inserting efficiency and flexibly controllable gene dosages. Additionally, simultaneous gene insertion at four genomic sites was achieved at 54.3% success rate while maintaining stable inheritance of exogenous sequences across multiple generations. The engineered strain could efficiently synthesize PHB from the fermentation of diverse organic wastes, with an efficiency comparable to those with overexpressed plasmid. When the mixture of seawater and molasses was used as the feedstock, it achieved a high PHB yield of 39.41 wt %. An extended application of the INTEGRET system for imparting the riboflavin production ability to the bacterium was also demonstrated. Our work presents a reliable and efficient genomic editing tool to facilitate the development of sustainable and environmentally benign biological platforms for converting biomass wastes into valuable chemicals.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"4233-4244"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142764602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Mitigating Winner-Take-All Resource Competition through Antithetic Control Mechanism.
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-12-06 DOI: 10.1021/acssynbio.4c00476
Suchana Chakravarty, Rishabh Guttal, Rong Zhang, Xiao-Jun Tian
{"title":"Mitigating Winner-Take-All Resource Competition through Antithetic Control Mechanism.","authors":"Suchana Chakravarty, Rishabh Guttal, Rong Zhang, Xiao-Jun Tian","doi":"10.1021/acssynbio.4c00476","DOIUrl":"10.1021/acssynbio.4c00476","url":null,"abstract":"<p><p>Competition among genes for limited transcriptional and translational resources impairs the functionality and modularity of synthetic gene circuits. Traditional control mechanisms, such as feedforward and negative feedback loops, have been proposed to alleviate these challenges, but they often focus on individual modules or inadvertently increase the burden on the system. In this study, we introduce three novel multimodule control strategies─local regulation, global regulation, and negatively competitive regulation (NCR)─that employ an antithetic regulatory mechanism to mitigate resource competition. Our systematic analysis reveals that while all three control mechanisms can alleviate resource competition to some extent, the NCR controller consistently outperforms both the global and local controllers. This superior performance stems from the unique architecture of the NCR controller, which is independent of specific parameter choices. Notably, the NCR controller not only facilitates the activation of less active modules through cross-activation mechanisms but also effectively utilizes the resource consumption within the controller itself. These findings emphasize the critical role of carefully designing the topology of multimodule controllers to ensure robust performance.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"4050-4060"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142783331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Stem Loop Mediated Transgene Modulation in Human T Cells.
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-12-06 DOI: 10.1021/acssynbio.4c00152
David Mai, Carly Harro, Aabir Sanyal, Philipp C Rommel, Neil C Sheppard, Carl H June
{"title":"Stem Loop Mediated Transgene Modulation in Human T Cells.","authors":"David Mai, Carly Harro, Aabir Sanyal, Philipp C Rommel, Neil C Sheppard, Carl H June","doi":"10.1021/acssynbio.4c00152","DOIUrl":"10.1021/acssynbio.4c00152","url":null,"abstract":"<p><p>Controlling gene expression is useful for many applications, but current methods often require external user inputs, such as the addition of a drug. We present an alternative approach using cell-autonomous triggers based on RNA stem loop structures in the 3' untranslated regions (UTRs) of mRNA. These stem loops are targeted by the RNA binding proteins Regnase-1 and Roquin-1, allowing us to program stimulation-induced transgene regulation in primary human T cells. By incorporating engineered stem loops into the 3' UTRs of transgenes, we achieved transgene repression through Regnase-1 and Roquin-1 activity, dynamic upregulation upon stimulation, and orthogonal tunability. To demonstrate the utility of this system, we employed it to modulate payloads in CAR-T cells. Our findings highlight the potential of leveraging endogenous regulatory machinery in T cells for transgene regulation and suggest RNA structure as a valuable layer for regulatory modulation.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"3897-3907"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142790502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Structure-Guided Engineering Unveils Deeper Substrate Channel in Processive Endoglucanase EG5C-1 Contributing to Enhanced Catalytic Efficiency and Processivity. 结构引导工程揭示了加工型内切葡聚糖酶 EG5C-1 中更深的底物通道,有助于提高催化效率和加工能力。
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-11-23 DOI: 10.1021/acssynbio.4c00562
Jialing Wang, Jie Chen, Kemin Lv, Zhen Gao, Jiahuang Li, Bin Wu, Bingfang He, Gerhard Schenk
{"title":"Structure-Guided Engineering Unveils Deeper Substrate Channel in Processive Endoglucanase EG5C-1 Contributing to Enhanced Catalytic Efficiency and Processivity.","authors":"Jialing Wang, Jie Chen, Kemin Lv, Zhen Gao, Jiahuang Li, Bin Wu, Bingfang He, Gerhard Schenk","doi":"10.1021/acssynbio.4c00562","DOIUrl":"10.1021/acssynbio.4c00562","url":null,"abstract":"<p><p>Processive endoglucanases have generated significant interest due to their bifunctionality in the degradation of cellulose and low product inhibition. However, enhancing their catalytic efficiency through engineering remains a formidable challenge. To address this bottleneck, our engineering efforts targeted loop regions located in the substrate channel of processive endoglucanase EG5C-1. Guided by a comparative analysis of characteristic structural features of the substrate channels in cellobiohydrolase, endoglucanase, and processive endoglucanase, a highly active triple mutant CM6 (N105H/T205S/D233L) was generated that had a 5.1- and 4.7-fold increase in catalytic efficiency toward soluble substrate carboxymethyl cellulose-Na and insoluble substrate phosphoric acid-swollen cellulose (PASC), compared with wild-type EG5C-1. Furthermore, this mutant exhibited greater processivity compared to EG5C-1. Molecular dynamics simulations unveiled that the mutations in the loop regions reshaped the substrate channel, leading to a deeper cleft, resembling the closed channel configuration of cellobiohydrolases. The increased compactness of the substrate channel induced changes in the substrate binding mode and substrate deformation, thereby enhancing both binding affinity and catalytic efficiency. Moreover, metadynamics simulations demonstrated that the processive velocity of cellulose chain through the binding channel in mutant CM6 surpassed that observed in EG5C-1.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"4131-4142"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142694875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
An Optimized Version of the Small Synthetic Genome (Mini-Synplastome) for Plastid Metabolic Engineering in Solanum tuberosum (Potato). 用于马铃薯质体代谢工程的小合成基因组(Mini-Synplastome)优化版。
IF 3.7 2区 生物学
ACS Synthetic Biology Pub Date : 2024-12-20 Epub Date: 2024-12-10 DOI: 10.1021/acssynbio.4c00724
Alessandro Occhialini, Gabriella King, Mohammad Majdi, Ivette A Fuentes Quispe, Jennifer M DeBruyn, Scott C Lenaghan
{"title":"An Optimized Version of the Small Synthetic Genome (Mini-Synplastome) for Plastid Metabolic Engineering in <i>Solanum tuberosum</i> (Potato).","authors":"Alessandro Occhialini, Gabriella King, Mohammad Majdi, Ivette A Fuentes Quispe, Jennifer M DeBruyn, Scott C Lenaghan","doi":"10.1021/acssynbio.4c00724","DOIUrl":"10.1021/acssynbio.4c00724","url":null,"abstract":"<p><p>Plastids represent promising targets in plant genetic engineering for many biotech applications, ranging from their use as bioreactors for the overproduction of valuable molecules to the installation of transgenes for improving plant traits. For over 30 years, routine methods of plastid transformation have relied on homologous recombination integrating vectors. However, nonintegrating episomal plasmids have recently received more attention as an innovative tool for the plastid genetic engineering of plant cells. One of these novel technologies is the mini-synplastome, an episomal plasmid with a chloroplast-specific origin of replication (<i>ori</i>) used to express transgenes in plastids. In order to improve episome sequence stability overtime by reducing the frequency of spurious recombination events, an optimized version of mini-synplastome (Gen3) was designed. The innovation in the Gen3 design was to substantially reduce the size of the plastomic sequence containing <i>oris</i> to include only domains involved in replication and to reduce the sequence homology of the whole episome with the endogenous plastome. In this work, we have demonstrated that Gen3 can be used to install a multigene pathway in <i>Solanum tuberosum</i> (potato) chloroplasts, and the episome is stable in a full-length circular form at high copy number throughout all plant developmental stages to anthesis in plants with normal phenotypic parameters. It is anticipated that in the next decade the mini-synplastome will be a valuable tool for installing complex genetic circuits in plastids.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"4245-4257"},"PeriodicalIF":3.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142826603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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