Synthetic biology (Oxford, England)最新文献

Going beyond size: exploring the metabolic burden in Pseudomonas putida during heterologous protein production. 超越大小：探索恶臭假单胞菌在异源蛋白生产过程中的代谢负担。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2026-02-12 eCollection Date: 2026-01-01 DOI: 10.1093/synbio/ysag004

Marleen Beentjes, Ana-Sofia Ortega-Arbulú, Carina Meiners, Andreas Kremling, Katharina Pflüger-Grau

{"title":"Going beyond size: exploring the metabolic burden in Pseudomonas putida during heterologous protein production.","authors":"Marleen Beentjes, Ana-Sofia Ortega-Arbulú, Carina Meiners, Andreas Kremling, Katharina Pflüger-Grau","doi":"10.1093/synbio/ysag004","DOIUrl":"https://doi.org/10.1093/synbio/ysag004","url":null,"abstract":"In biotechnological applications, it is often necessary to introduce genes or entire pathways into a host cell, which can create a significant metabolic burden on the host, limiting productivity. In this study, we systematically investigated the physiological stress responses of Pseudomonas putida during heterologous protein production using a modular monitoring system consisting of a plasmid encoding a heterologous protein fused to eGFP and a chromosomally integrated capacity reporter. Our findings reveal that translation is the main bottleneck, with translational capacity becoming saturated under high expression loads. While increasing the strength of the ribosome binding site improved protein production for non-burdensome proteins, this effect was not observed for larger fusion proteins. Variations in fusion protein size suggested that translational demand, rather than the overall mass of protein produced, determines metabolic burden. We further evaluated how resource availability affects protein expression by modifying the metabolic regime or supplementing with amino acids. While the carbon source affected cellular capacity, it did not significantly alter heterologous protein production. Amino acid supplementation alleviated the growth defects of MBPeGFP-producing cells and modestly improved protein production rates. Together, these findings emphasize that metabolic burden is influenced not only by the size of the produced protein but also by transcript architecture, resource allocation, and the physiological state of the host. Therefore, successful optimization of heterologous protein production requires a holistic approach integrating construct design with host physiology and cultivation strategies.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"11 1","pages":"ysag004"},"PeriodicalIF":2.5,"publicationDate":"2026-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12998221/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147488974","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}

引用次数: 0

A protein-fragment complementation assay to quantify synthetic protein scaffold efficiency. 定量合成蛋白支架效率的蛋白质片段互补试验。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2026-01-28 eCollection Date: 2026-01-01 DOI: 10.1093/synbio/ysag003

Pascale Lemieux, Alexandre K Dubé, Christian R Landry

{"title":"A protein-fragment complementation assay to quantify synthetic protein scaffold efficiency.","authors":"Pascale Lemieux, Alexandre K Dubé, Christian R Landry","doi":"10.1093/synbio/ysag003","DOIUrl":"https://doi.org/10.1093/synbio/ysag003","url":null,"abstract":"Scaffolds are powerful tools in synthetic biology used for various applications, from increasing yield to optimizing signalling specificity. Protein scaffolds can be built by fusing peptide binding domains (PBD) and attaching the peptide they bind to enzymes, inducing spatial proximity. Only a few PBD-peptide combinations have been tested in this context, and no combination produced a high yield in yeast, an important chassis in biotechnology. Therefore, there is a need for more exploration of PBD-peptide pairs to be used in this model. Scaffold characterization is challenging because it is often dependent on a model pathway with an output that is difficult to measure quantitatively. Here, we use the dihydrofolate reductase protein-fragment complementation assay (DHFR PCA) to study scaffolding efficiency in yeast, which allows to couple scaffolding efficiency with growth rate. First, we characterize the strength of PBD-peptide interactions (PPI) and the binding availability of the PBDs and peptides. Then, we test different scaffold architectures and expression levels to quantify the simultaneous binding of peptide pairs to the scaffold. We show that PPI strength of the weakest binding PBD-peptide pair is critical for scaffolding efficiency and that PPI strength is limited by low binding availability of some domains and peptides in vivo. Also, we find that slight architectural variations and expression levels have a significant impact on scaffolding efficiency detected by DHFR PCA. Finally, we used DHFR PCA approaches to characterize novel PBD-peptide pairs and we identified pairs to expand the sequence toolbox for scaffold design in yeast through DHFR PCA easy-to-read signal.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"11 1","pages":"ysag003"},"PeriodicalIF":2.5,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12981675/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147470491","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}

引用次数: 0

EASY-C: Extraction and Analysis of Small Yeast Chromosomes-A rapid and universal platform for recovering artificial mini-chromosomes from synthetic Sc2.0 yeast and large plasmids from Saccharomyces cerevisiae and nonconventional yeast species. EASY-C：小酵母染色体的提取和分析-一个快速和通用的平台，用于从合成的Sc2.0酵母和从酿酒酵母和非常规酵母中提取大质粒的人工小染色体。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2026-01-21 eCollection Date: 2026-01-01 DOI: 10.1093/synbio/ysag002

Reem Swidah, Marco Monti, Daniela Delneri

{"title":"EASY-C: Extraction and Analysis of Small Yeast Chromosomes-A rapid and universal platform for recovering artificial mini-chromosomes from synthetic Sc2.0 yeast and large plasmids from Saccharomyces cerevisiae and nonconventional yeast species.","authors":"Reem Swidah, Marco Monti, Daniela Delneri","doi":"10.1093/synbio/ysag002","DOIUrl":"https://doi.org/10.1093/synbio/ysag002","url":null,"abstract":"The protocol for Extraction and Analysis of Small Yeast-Chromosomes (EASY-C) is a transformative, rapid, cost-effective, and user-friendly method designed for the efficient isolation of artificial synthetic mini-chromosomes (~42-52 kb) and large plasmids (~12 kb) from Saccharomyces cerevisiae wild-type and synthetic yeast strains (Sc2.0), and a range of nonconventional yeast (NCY) species. In this two-step workflow, the DNA from yeast is first extracted and transferred into bacteria, and then the circular DNA is recovered from the bacteria and subjected to downstream analysis, including long-read sequencing. The EASY-C protocol operates at small volumes (~1 mL) and requires less than 2.5 hours, allowing the use of standard commercial plasmid purification kits for bacterial plasmids. Under the tested conditions, the EASY-C methodology yielded clean DNA that could be digested and linearized prior to sequencing, resulting in a higher number of high-quality reads (~2000). The EASY-C protocol worked successfully for the extraction of a variety of constructs, including low-copy centromeric vectors (CEN/ARS), high-copy plasmids (pan/ARS), and artificial mini-chromosomes harbouring (CEN/ARS). It is also applicable to a variety of yeast species, including NCY such as Starmerella sp., Maudiozyma sp., and Kazachstania sp. Thanks to its precision, robustness, and simplicity, EASY-C equips researchers with a powerful, time-saving tool and cost-effective approach to accelerate the validation of a wide array of synthetic genetic and metabolic constructs engineered in vivo across diverse yeast species.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"11 1","pages":"ysag002"},"PeriodicalIF":2.5,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922780/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147273554","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}

引用次数: 0

Development and characterization of pNarsenic: a naringenin-inducible biosensor for arsenic in Escherichia coli. 柚皮素诱导的大肠杆菌砷生物传感器pNarsenic的研制与表征。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2026-01-10 eCollection Date: 2026-01-01 DOI: 10.1093/synbio/ysag001

Vincent Crabbe, Ezgi Unal, Stijn De Graeve, Daniel G Guerra, Tom Peeters, Sophie de Buyl, Eveline Peeters, Indra Bervoets

{"title":"Development and characterization of pNarsenic: a naringenin-inducible biosensor for arsenic in Escherichia coli.","authors":"Vincent Crabbe, Ezgi Unal, Stijn De Graeve, Daniel G Guerra, Tom Peeters, Sophie de Buyl, Eveline Peeters, Indra Bervoets","doi":"10.1093/synbio/ysag001","DOIUrl":"https://doi.org/10.1093/synbio/ysag001","url":null,"abstract":"Whole-cell biosensors detecting the heavy metal arsenic have been widely studied for their potential in environmental monitoring. And while inducible biosensors have been shown to be an effective tool to tune the operational range, a thoroughly characterized inducible biosensor is currently lacking. Here, we present an Escherichia coli biosensor for arsenic in which the transcription factor (TF) gene arsR is inducible by naringenin, a plant-derived secondary metabolite. Increasing the naringenin concentration reduced the basal output while increasing both the dynamic range and sensing threshold of the biosensor dose-response curve, but the operational range appeared constrained by a fixed upper limit. Comparison with a previously published phenomenological model revealed good overall agreement between experimental data and model predictions, except for the behaviour of the maximum output and threshold. This work expands the biosensor toolbox with a profoundly characterized arsenic biosensor and raises a potential practical limit to dose-response curve engineering by tuning TF expression alone.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"11 1","pages":"ysag001"},"PeriodicalIF":2.5,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12840587/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146095136","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}

引用次数: 0

RAPID-DASH: fast and efficient assembly of guide RNA arrays for multiplexed CRISPR-Cas9 applications. RAPID-DASH：用于多路CRISPR-Cas9应用的快速高效的向导RNA阵列组装。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2025-12-18 eCollection Date: 2026-01-01 DOI: 10.1093/synbio/ysaf020

Asfar Lathif Salaudeen, Nicholas Mateyko, Carl G de Boer

引用次数: 0

Emerging trends in genome integration tools for precision engineering of diverse bacterial species. 不同细菌物种精密工程的基因组整合工具的新趋势。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2025-12-11 eCollection Date: 2025-01-01 DOI: 10.1093/synbio/ysaf019

Riesa K W Rohmat, Thea C T Irvine, Shivang Hina-Nilesh Joshi, Andrew M Bailey, Christopher Jenkins, David Ulaeto, Pierre Buscaill, Thomas E Gorochowski

{"title":"Emerging trends in genome integration tools for precision engineering of diverse bacterial species.","authors":"Riesa K W Rohmat, Thea C T Irvine, Shivang Hina-Nilesh Joshi, Andrew M Bailey, Christopher Jenkins, David Ulaeto, Pierre Buscaill, Thomas E Gorochowski","doi":"10.1093/synbio/ysaf019","DOIUrl":"10.1093/synbio/ysaf019","url":null,"abstract":"The ability to precisely insert DNA payloads into a genome enables the comprehensive engineering of cellular phenotypes and the creation of new biotechnologies. To achieve such modifications, the most widely used techniques rely on a host cell's native DNA repair mechanisms like homologous recombination, which hampers their broader use in organisms lacking these capabilities. Here, we explore the current landscape of genome integration systems with a particular focus on those that function in bacteria and are precise, self-contained, and portable, placing minimal requirements on the host cell. Through a historical analysis, we observe long-term use of recombineering technologies, a recent rise in the use of CRISPR-guided systems that consist of associated integrase machinery, and growing efforts to modify non-model organisms. Looking forward, we highlight some of the remaining challenges and how synthetic genomics may offer a way to create bacterial strains optimized for extensive long-term modification. As the field of synthetic biology sets its sights on real-world impact, the effective engineering of genomes will be critical to shaping the robust phenotypes that applications demand.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"10 1","pages":"ysaf019"},"PeriodicalIF":2.5,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12765450/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907441","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}

引用次数: 0

Expanding the Yeast MoClo Toolkit: gene expression control parts for Saccharomyces cerevisiae tested in industrially relevant conditions. 扩展酵母MoClo工具包：在工业相关条件下测试的酿酒酵母基因表达控制部分。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2025-12-04 eCollection Date: 2025-01-01 DOI: 10.1093/synbio/ysaf018

Elin Blick, Yvonne Nygård

{"title":"Expanding the Yeast MoClo Toolkit: gene expression control parts for Saccharomyces cerevisiae tested in industrially relevant conditions.","authors":"Elin Blick, Yvonne Nygård","doi":"10.1093/synbio/ysaf018","DOIUrl":"10.1093/synbio/ysaf018","url":null,"abstract":"Fine-tuning of gene expression is often required to achieve competitive production levels in microbial cell factories. Several orthogonal expression systems based on heterologous regulatory parts have been developed for Saccharomyces cerevisiae. In laboratory conditions the systems demonstrate predictable results, but few expression systems have been tested in industrial conditions. Here, a new expression system based on the bacterial gene cusR was developed for S. cerevisiae, and two previous developed systems, the strong Bm3R1-based system and the quinic acid inducible Q-system, were adapted for compatibility with the Yeast MoClo Toolkit. The bacterial transcription factors CusR and Bm3R1 acted as DNA binding domains, and fused to a viral activation domain, they functioned as transcriptional activators. The Q-system is originally from Neurospora crassa and consists of a transcriptional repressor, QS, which in the absence of quinic acid blocks the activity of a transcriptional activator, QF2. Quinic acid binds to QS, inhibiting QS from blocking the activity of QF2 in a dose-dependent manner. The gene expression systems were assessed in industrially relevant conditions, proving a predictable performance at low pH. The performance of the constitutive systems was predictable also at high temperature and in a synthetic lignocellulosic hydrolysate medium. Altogether, the MoClo-compatible expression systems enable fast construction of fine-tuned production pathways for S. cerevisiae cell factories used for industrial applications.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"10 1","pages":"ysaf018"},"PeriodicalIF":2.5,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12721388/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145822353","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}

引用次数: 0

Engineering artificial 5' regulatory sequences for thermostable protein expression in the extremophile Thermus thermophilus. 在嗜热嗜热菌中表达热稳定蛋白的工程人工5′调控序列。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2025-12-04 eCollection Date: 2025-01-01 DOI: 10.1093/synbio/ysaf016

Che Fai Alex Wong, Shizhe Zhang, Lisa Tietze, Gurvinder Singh Dahiya, Rahmi Lale

{"title":"Engineering artificial 5' regulatory sequences for thermostable protein expression in the extremophile Thermus thermophilus.","authors":"Che Fai Alex Wong, Shizhe Zhang, Lisa Tietze, Gurvinder Singh Dahiya, Rahmi Lale","doi":"10.1093/synbio/ysaf016","DOIUrl":"10.1093/synbio/ysaf016","url":null,"abstract":"The utilization of biocatalysts in biotechnological applications often necessitates their heterologous expression in suitable host organisms. However, the range of standardized microbial hosts for recombinant protein production remains limited, with most being mesophilic and suboptimal for certain protein types. Although the thermophilic bacterium Thermus thermophilus has long been established as a valuable extremophile host, thanks to its high-temperature tolerance, robust growth, and extensively characterized proteome, its genetic toolkit has predominantly depended on a limited set of native promoters. To overcome this bottleneck, we have expanded the available regulatory repertoire in T. thermophilus by developing novel artificial 5[Formula: see text] regulatory sequences (ARESs). In this study, we applied our Gene Expression Engineering platform to engineer 53 artificial ARES in T. thermophilus. These ARES, which comprise both promoter and 5[Formula: see text] untranslated regions, were functionally characterized in both T. thermophilus and Escherichia coli, revealing distinct host-specific expression patterns. Furthermore, we demonstrated the utility of these ARES by achieving high-level expression of thermostable proteins, including [Formula: see text]-galactosidase, a superfolder citrine fluorescent protein, and phytoene synthase. A bioinformatic analysis of the novel sequences has also been carried out indicating that the ARES possess markedly lower Guanine (G) and Cytosine (GC) content compared to native promoters. This study contributes to expanding the genetic toolkit for recombinant protein production by providing a set of functionally validated ARES, enhancing the versatility of T. thermophilus as a synthetic biology chassis for thermostable protein expression.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"10 1","pages":"ysaf016"},"PeriodicalIF":2.5,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12704435/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145770038","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}

引用次数: 0

Harnessing mass spectrometry-based proteomics for continuous directed evolution. 利用基于质谱的蛋白质组学进行连续定向进化。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2025-12-04 eCollection Date: 2025-01-01 DOI: 10.1093/synbio/ysaf017

Katharina Belt, David Obe, Mark A Wilson, A Harvey Millar, Ulschan Bathe

{"title":"Harnessing mass spectrometry-based proteomics for continuous directed evolution.","authors":"Katharina Belt, David Obe, Mark A Wilson, A Harvey Millar, Ulschan Bathe","doi":"10.1093/synbio/ysaf017","DOIUrl":"10.1093/synbio/ysaf017","url":null,"abstract":"Continuous directed evolution is a powerful Synthetic Biology tool to engineer proteins with desired functions in vivo. Mimicking natural evolution, it involves repeated cycles of high-frequency mutagenesis, selection, and replication within platform cells, where the function of the target gene is tightly linked to the host cell's fitness. However, cells might escape the selection pressure due to the inherent flexibility of their metabolism, which allows for adaptation. Whole-proteome analysis as well as targeted proteomics offer valuable insights into global and specific cellular changes. They can identify modifications in the target protein and its interactors to help understand its evolution and network integration. Using the continuous evolution of the Arabidopsis thaliana methionine synthases AtMS1 and AtMS2 as an example, we show how mass spectrometry-based proteomics was able to assess the abundance of target enzymes, identify flaws in population construction, measure methionine metabolic adaptation, and allow informed decision-making in the evolution campaign.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"10 1","pages":"ysaf017"},"PeriodicalIF":2.5,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12765272/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907473","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}

引用次数: 0

GoldenBraid2.0 E. coli: a comprehensive and characterized toolkit for enterics. GoldenBraid2.0大肠杆菌：一个全面和特征的肠道工具包。

IF 2.5

Synthetic biology (Oxford, England) Pub Date : 2025-08-14 eCollection Date: 2025-01-01 DOI: 10.1093/synbio/ysaf015

Matthew B Cooke, Kobie T Welch, Laura D Ramirez, Alice X Wen, David C Marciano, Christophe Herman

{"title":"GoldenBraid2.0 E. coli: a comprehensive and characterized toolkit for enterics.","authors":"Matthew B Cooke, Kobie T Welch, Laura D Ramirez, Alice X Wen, David C Marciano, Christophe Herman","doi":"10.1093/synbio/ysaf015","DOIUrl":"10.1093/synbio/ysaf015","url":null,"abstract":"Modular cloning systems streamline laboratory workflows by consolidating genetic 'parts' into reusable and modular collections, enabling researchers to fast-track strain construction. The GoldenBraid 2.0 modular cloning system utilizes the cutting property of type IIS restriction enzymes to create defined genetic 'grammars', which facilitate the reuse of standardized genetic parts and assembly of genetic parts in the right order. Here, we present a GoldenBraid 2.0 toolkit of genetic parts designed to accelerate cloning in the model bacterium Escherichia coli. This toolkit features 478 pre-made parts for gene expression and protein tagging as well as strains to expedite cloning and strain construction, enabling researchers to quickly generate functional plasmid-borne or chromosome-integrated expression constructs. In addition, we provide a complete laboratory manual with overviews of common reagent recipes, E. coli protocols, and community resources to promote toolkit utilization. By streamlining the assembly process, this resource will reduce the financial and temporal burdens of cloning and strain building in many laboratory settings.","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"10 1","pages":"ysaf015"},"PeriodicalIF":2.5,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12415853/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145031464","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}

引用次数: 0