Synthetic and Systems Biotechnology最新文献

{"title":"Integration of metabolic and evolutionary processes to construct efficient xylose-utilizing strain of Zymomonas mobilis for lignocellulosic ethanol production","authors":"Jiyun Lou ,&nbsp;Xia Wang ,&nbsp;Runxia Li ,&nbsp;Jun Yao ,&nbsp;Junpeng Hu ,&nbsp;Xianwu Qin ,&nbsp;Mingjie Jin ,&nbsp;Shihui Yang","doi":"10.1016/j.synbio.2025.04.009","DOIUrl":"10.1016/j.synbio.2025.04.009","url":null,"abstract":"<div><div>Xylose is a major component of lignocellulose, and it is crucial to enable microorganisms the capability of efficient xylose utilization for economical lignocellulosic biochemical production. <em>Zymomonas mobilis</em> is a natural ethanologenic bacterium with xylose-utilizing recombinant strains engineered for commercial lignocellulosic ethanol production. However, low efficiency of xylose utilization is still the major hurdle for lignocellulosic biochemical production, especially in the lignocellulosic hydrolysate containing inhibitory compounds and mixed sugars of glucose and xylose. In this study, xylose-utilization capability of a xylose-utilizing recombinant strain 8b-S38 of <em>Z. mobilis</em> was further improved by evaluating the effects of different xylose metabolic pathways, xylose isomerases, and xylose transporters. The results demonstrated that xylose isomerase pathway is still the most efficient one in <em>Z. mobilis</em> among the xylose metabolic pathways examined, and the introduction of xylose isomerase (XI) such as StrXI and PanXI to increase its copy numbers can improve xylose utilization. In addition, the introduction of effective xylose transporter such as high-affinity xylose transporter XylFGH and the glucose facilitator mutant Glf^{A165M−K458I} promoted xylose transportation and subsequent utilization. A recombinant strain S67SPGlf^m was constructed combining rational engineering of xylose isomerases and transporters and semi-rational adaptation, which exhibited enhanced xylose utilization capability in lignocellulosic hydrolysate. This study illustrated that the construction of robust and efficient industrial strains with complex phenotypes requires a combination of different strategies such as metabolic engineering and adaptive laboratory evolution, which also provided biological parts such as xylose isomerases and xylose transporters to help design and construct microbial cell factories for efficient xylose utilization in the future.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 858-867"},"PeriodicalIF":4.4,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869540","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}

{"title":"Nonglycosidic C–O bond formation catalyzed by a bifunctional pseudoglycosyltransferase ValL","authors":"Ziyue Guo ,&nbsp;Xin Zhang ,&nbsp;Lin Zhou ,&nbsp;Qungang Huang ,&nbsp;Qianjin Kang ,&nbsp;Linquan Bai","doi":"10.1016/j.synbio.2025.04.007","DOIUrl":"10.1016/j.synbio.2025.04.007","url":null,"abstract":"<div><div>The C₇N antibiotic validamycin A is an antifungal agent widely used as a crop protectant. It comprises a validoxylamine A unit linked to a glucose moiety, which is formed through a nonglycosidic <em>C</em><em>–</em><em>N</em> bond connecting a valienol moiety and a validamine moiety, a reaction catalyzed by the pseudoglycosyltransferase ValL. In this study, we analyzed the chemical composition of validamycins in <em>Streptomyces hygroscopicus</em> var. <em>jinggangensis</em> TL01. A series of novel oxygen-bridged analogues, namely, validenomycin, validomycin, and 1,1′-bis-valienol, were identified in the culture supernatants, and their chemical structures were elucidated using a combination of one- and two-dimensional nuclear magnetic resonance and mass spectrometry. Gene disruption and complementation experiments revealed that <em>valL</em> is essential for the biosynthesis of these new oxygen-bridged analogues of validamycins. Biochemical assays further demonstrated that ValL catalyzed the <em>C–O</em> bond formation between GDP-valienol and valienol-7-phosphate, producing 1,1′-bis-valienol-7-phosphate, which was subsequently dephosphorylated by ValO and glycosylated by ValG to yield validenomycin. Collectively, our findings revealed the unique ability of ValL to catalyze nonglycosidic <em>C–O</em> coupling, potentially enabling the generation of various chemical scaffolds for C₇N family antibiotics.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 846-857"},"PeriodicalIF":4.4,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143864111","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}

{"title":"A systematic study of regulating inorganic polyphosphates production in Saccharomyces cerevisiae","authors":"Zipeng Chen ,&nbsp;Yanling Wu ,&nbsp;Lingfeng Qin ,&nbsp;Chen Wang ,&nbsp;Zhixin Li ,&nbsp;Xiaozhou Luo ,&nbsp;Wei Wei ,&nbsp;Jing Zhao","doi":"10.1016/j.synbio.2025.04.004","DOIUrl":"10.1016/j.synbio.2025.04.004","url":null,"abstract":"<div><div>Inorganic polyphosphate (polyP), a linear polymer of orthophosphate residues, plays critical roles in diverse biological processes spanning blood coagulation, immunomodulation, and post-translational protein modifications in eukaryotes. Notably, long-chain polyP (&gt;100 phosphate units) exhibits distinct biological functionalities compared to shorter-chain counterparts. While <em>Saccharomyces cerevisiae</em> serves as a promising microbial platform for polyP biosynthesis, the genetic regulatory mechanisms underlying polyP metabolism remain poorly elucidated. Here, we systematically investigated the genetic determinants governing intracellular polyP levels and chain length dynamics in yeast. Through screening a library of 55 single-gene knockout strains, we identified six mutants (<em>Δddp1</em>, <em>Δvip1</em>, <em>Δppn1</em>, <em>Δppn2</em>, <em>Δecm33</em>, and <em>Δccr4</em>) exhibiting elevated polyP accumulation, whereas deletions of <em>vtc1</em>, <em>kcs1</em>, <em>vma22</em>, <em>vma5</em>, <em>pho85</em>, <em>vtc4</em>, <em>vma2</em>, <em>vma3</em>, <em>ecm14</em>, and <em>vph2</em> resulted in near-complete polyP depletion. Subsequent combinatorial deletions in the <em>Δppn1</em> background revealed that the <em>Δppn1Δvip1</em> double mutant achieved synergistic enhancement in both polyP concentration (53.01 mg-P/g-DCW) and chain length, attributable to increased ATP availability and reduced polyphosphatase activity. Leveraging CRISPR/Cas9-mediated overexpression in <em>Δppn1Δvip1</em>, we engineered strain PP2 (<em>vtc4</em> overexpression), which demonstrated a 2-fold increase in polyP yield (62.6 mg-P/g-DCW) relative to wild-type BY4741, with predominant synthesis of long-chain species. Mechanistically, qRT-PCR analysis confirmed that PP2 exhibited 46-fold up-regulation of <em>vtc4</em> coupled with down-regulation of polyphosphatases encoding genes, <em>ppn2</em>, <em>ddp1</em>, and <em>ppx1</em>. This study performed a systematic study of regulating inorganic polyphosphates production in yeast and provides a synthetic biology strategy to engineer high-yield polyP-producing strains, advancing both fundamental understanding and biotechnological applications.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 816-826"},"PeriodicalIF":4.4,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848255","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}

{"title":"Enhanced l-serine synthesis in Corynebacterium glutamicum by exporter engineering and Bayesian optimization of the medium composition","authors":"Yifan Huang ,&nbsp;Yujie Gao ,&nbsp;Yamin Huang ,&nbsp;Xiaogang Wang ,&nbsp;Meijuan Xu ,&nbsp;Guoqiang Xu ,&nbsp;Xiaojuan Zhang ,&nbsp;Hui Li ,&nbsp;Jinsong Shi ,&nbsp;Zhenghong Xu ,&nbsp;Xiaomei Zhang","doi":"10.1016/j.synbio.2025.04.003","DOIUrl":"10.1016/j.synbio.2025.04.003","url":null,"abstract":"<div><div><span>l</span>-serine is a versatile, high value-added amino acid, widely used in food, medicine and cosmetics. However, the low titer of <span>l</span>-serine has limited its industrial production. In this study, a cell factory without plasmid for efficient production of <span>l</span>-serine was constructed based on transport engineering. Firstly, the effects of <span>l</span>-serine exporter SerE overexpression and deletion on the cell growth and <span>l</span>-serine titer were investigated in <em>Corynebacterium glutamicum</em> (<em>C. glutamicum</em>) A36, overexpression of <em>s</em><em>erE</em> using a plasmid led to a 15.1% increase in <span>l</span>-serine titer but also caused a 15.1% decrease in cell growth. Subsequently, to increase the export capacity of SerE, we conducted semi-rational design and bioinformatics analysis, combined with alanine mutation and site-specific saturation mutation. The mutant E277K was obtained and exhibited a 53.2% higher export capacity compared to wild-type SerE, resulting in <span>l</span>-serine titer increased by 39.6%. Structural analysis and molecular dynamics simulations were performed to elucidate the mechanism. The results showed that the mutation shortened the hydrogen bond distance between the exporter and <span>l</span>-serine, enhanced complex stability, and reduced the binding energy. Finally, Bayesian optimization was employed to further improve <span>l</span>-serine titer of the mutant strain C-E277K. Under the optimized conditions, 47.77 g/L <span>l</span>-serine was achieved in a 5-L bioreactor, representing the highest reported titer for <em>C. glutamicum</em> to date. This study provides a basis for the transformation of <span>l</span>-serine export pathway and offers a new strategy for increasing <span>l</span>-serine titer.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 835-845"},"PeriodicalIF":4.4,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143850456","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}

{"title":"dTSR enables efficient improvement of heterologous production of spinosad in Saccharopolyspora erythraea","authors":"Weijun He ,&nbsp;Jin Lü ,&nbsp;Lulu Bai ,&nbsp;Zixin Deng ,&nbsp;Meifeng Tao","doi":"10.1016/j.synbio.2025.02.003","DOIUrl":"10.1016/j.synbio.2025.02.003","url":null,"abstract":"<div><div>Spinosad (spinosyns A and D) is a highly effective and environmentally friendly insecticide widely used for pest control. However, the difficulty of genetic manipulation in the original strain, <em>Saccharopolyspora spinosa</em>, has hindered improvements in fermentation yields using synthetic biology methods. Additionally, there is a lack of simple and effective methods for enhancing the production of polyketide natural products derived from slow-growing rare actinomycetes. In this study, we developed a doubly transposition and site-specific recombination (dTSR) approach to insert bacterial attachment sites (<em>attB</em>) and two copies of spinosad biosynthetic gene cluster (<em>spn</em> BGC) into various chromosomal locations of <em>Saccharopolyspora erythraea</em>, thereby generating heterologous production strains for screening of spinosyn producers with improved yields. Engineered strains from the first round of TSR breeding produced spinosad at levels ranging from 5.6 to 30.5 mg/L. The second round of TSR breeding produced engineered strains with increased yields, with the highest spinosad production reaching 137.1 ± 10.9 mg/L. These results indicated that (1) the dTSR approach could efficiently generate initial heterologous strains with significantly improved spinosad production, and (2) the dTSR approach enabled random integration of a second copy of <em>spn</em> BGC into various chromosomal locations in <em>Sac. erythraea</em>, thereby further increasing heterologous spinosad production to high levels. This study provides a simple, rational, and efficient approach to improve the heterologous production of polyketide natural products in rare actinomycetes.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 868-875"},"PeriodicalIF":4.4,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143877049","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}

{"title":"Programming anti-ribozymes to sense trigger RNAs for modulating gene expression in mammalian cells","authors":"Wenhui Zhang ,&nbsp;Shi Zhao ,&nbsp;Mengyuan Wang ,&nbsp;Chunbo Lou ,&nbsp;Yanhui Xiang ,&nbsp;Qiong Wu","doi":"10.1016/j.synbio.2025.03.011","DOIUrl":"10.1016/j.synbio.2025.03.011","url":null,"abstract":"<div><div>Synthetic RNA-based switches provide distinctive merits in modulating gene expression. Simple and flexible RNA-based switches are crucial for advancing the field of gene regulation, paving the way for innovative tools that can sense and manipulate cellular processes. In this research, we have developed programmable ribozymes that are capable of suppressing gene expression in response to specific, endogenously expressed trigger RNAs. We engineer ribozymes by introducing upstream antisense sequences (anti-ribozymes) to inhibit the self-cleaving activity of the hammerhead ribozyme and open the expression of the target gene. The trigger RNA is designed to recognize and bind to complementary sequences within the anti-ribozymes, thereby inhibiting their ability to direct protein synthesis. The anti-ribozyme performance is optimized by regulating the essential sequence modules that play a crucial role in determining the specificity and efficiency of the anti-ribozyme's interaction with its trigger RNA. By applying this switch mechanism to various ribozyme designs, we have shown that it is possible to achieve control over gene expression across a wide range of trigger RNAs. By exploiting these programmable anti-ribozymes, we aim to create a powerful tool for controlling gene expression in mammalian cells, which could have important implications for basic research, disease diagnosis, and therapeutic interventions.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 827-834"},"PeriodicalIF":4.4,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848256","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}

{"title":"A new paradigm for the regulation of A40926B0 biosynthesis","authors":"Yan-Qiu Liu ,&nbsp;Yi-Lei Zheng ,&nbsp;Ye Xu ,&nbsp;Xue-Yan Liu ,&nbsp;Tian-Yu Xia ,&nbsp;Qing-Wei Zhao ,&nbsp;Yong-Quan Li","doi":"10.1016/j.synbio.2025.03.012","DOIUrl":"10.1016/j.synbio.2025.03.012","url":null,"abstract":"<div><div>Dalbavancin is a novel glycopeptide antibiotic with activity against a broad range of Gram-positive bacteria, including methicillin-resistant <em>Staphylococcus aureus</em> (MRSA). A40926B0 is the direct precursor of dalbavancin, but the regulatory mechanisms underlying its biosynthesis are not well understood. Additionally, the presence of seven homologs leads to significant metabolic burden, affecting both production and purity of A40926B0. To further reveal the transcriptional regulatory mechanism of A40926B0 biosynthesis in <em>N. gerenzanensis</em> L70, this study employed multiple strategies to explore the regulatory network of its biosynthesis from both intracluster and extracluster aspects. WblA regulates gene expression within and outside the biosynthetic gene cluster (BGC), impacting multiple biosynthetic pathways, and Dbv3, a key regulator in the A40926B0 cluster, positively influences biosynthesis. Using a bottom-up (DNA to protein) regulator mining strategy with the key intra-cluster regulator Dbv3, it was determined that GlnR is also involved in the regulation of secondary metabolism, while BkdR regulates precursor supply. By constructing the combination of GlnR, BkdR and Dbv3 together with the WblA deletion, the regulatory network of A40926B0 was reconstructed, resulting in a 92 % improvement in purity of A40926B0. The objective of this study is to elucidate the regulatory mechanisms governing A40926B0 biosynthesis by constructing a comprehensive, multidimensional model of its regulatory network. The findings of this study serve to enhance our comprehension of A40926B0 biosynthesis and furnish insights into broader strategies for enhancing the production of other natural products and secondary metabolites in industrial microbiology.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 794-806"},"PeriodicalIF":4.4,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143843693","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}

{"title":"Efficient production of 2′-fucosyllactose in Pichia pastoris through metabolic engineering and constructing an orthogonal energy supply system","authors":"Yi Li ,&nbsp;Xiang Wang ,&nbsp;Kaidi Chen ,&nbsp;Zhoukang Zhuang ,&nbsp;Hongting Tang ,&nbsp;Tao Yu ,&nbsp;Wenbing Cao","doi":"10.1016/j.synbio.2025.04.002","DOIUrl":"10.1016/j.synbio.2025.04.002","url":null,"abstract":"<div><div>2′-fucosyllactose (2′-FL) holds significant role in the infants' nutrition. While microbial production of 2′-FL has predominantly utilized <em>Escherichia coli</em> and <em>Saccharomyces cerevisiae</em>, the potential of <em>Pichia pastoris</em>, renowned for its robust NADPH regeneration capability, remains underexplored. Herein, we systematically engineered the metabolism of <em>P. pastoris</em> to develop an efficient 2′-FL-producing cell factory. We first constructed the <em>de novo</em> biosynthesis pathway for 2′-FL in <em>P. pastoris</em>, achieving an initial titer of 0.143 g/L. By optimizing enzyme selection and solubility of α-1,2-fucosyltransferase (FutC), 2′-FL production was enhanced by nearly ten folds. Subsequently, engineering NADPH supply further increased the 2′-FL production by 170 %. Furthermore, we enhanced energy supply by incorporating an orthogonal energy module based on the methanol dissimilation pathway and increasing GTP availability, resulting in a 32 % improvement in 2′-FL production. Finally, through the optimization of fermentation condition, we realized the production titer of 2′-FL to 3.50 g/L in shake-flask, representing the highest titer in <em>P. pastoris</em>. These findings highlight the potential of <em>P. pastoris</em> as a chassis to produce chemicals by providing abundant NADPH and utilizing methanol as co-substrate to supply sufficient energy.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 807-815"},"PeriodicalIF":4.4,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143843694","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}

{"title":"MTD: A cloud-based omics database and interactive platform for Myceliophthora thermophila","authors":"Jiacheng Dong ,&nbsp;Zhitao Mao ,&nbsp;Haoran Li ,&nbsp;Ruoyu Wang ,&nbsp;Yutao Wang ,&nbsp;Haokai Jia ,&nbsp;Jingen Li ,&nbsp;Qian Liu ,&nbsp;Chenglin Zhang ,&nbsp;Xiaoping Liao ,&nbsp;Defei Liu ,&nbsp;Hongwu Ma ,&nbsp;Chaoguang Tian","doi":"10.1016/j.synbio.2025.04.001","DOIUrl":"10.1016/j.synbio.2025.04.001","url":null,"abstract":"<div><div>Nowadays, biological databases are playing an increasingly critical role in biological research. <em>Myceliophthora thermophila</em> is an excellent thermophilic fungal chassis for industrial enzyme production and plant biomass-based chemical synthesis. The lack of a dedicated public database has made access to and reanalysis of <em>M. thermophila</em> data difficult. To bridge this gap, we developed MTD (<span><span>https://mtd.biodesign.ac.cn/</span><svg><path></path></svg></span>), a cloud-based omics database and interactive platform for <em>M. thermophila</em>. MTD integrates comprehensive genome annotations, sequence-based predictions, transcriptome data, curated experimental descriptions, and bioinformatics analysis tools, offering a comprehensive, one-stop solution with a ‘top-down’ search strategy to streamline <em>M. thermophila</em> research. The platform supports data reproduction, rapid querying, and in-depth mining of existing transcriptome datasets. Based on analyses using data and tools in MTD, we identified shifts in metabolic allocation in a glucoamylase hyperproduction strain of <em>M. thermophila</em>, highlighting changes in fatty acid biosynthesis and amino acids biosynthesis pathways, which provide new insights into the underlying phenotypic alterations. As a pioneering resource, MTD marks a key advancement in <em>M. thermophila</em> research and sets the model for developing similar databases for other species.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 783-793"},"PeriodicalIF":4.4,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143829280","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}

{"title":"Development of ultrasound-visualized tumor-targeting engineered bacteria for precise tumor therapy","authors":"Li Qu,&nbsp;Zhou Chi,&nbsp;Zhen-Ping Zou,&nbsp;Ying Zhou,&nbsp;Bang-Ce Ye","doi":"10.1016/j.synbio.2025.03.009","DOIUrl":"10.1016/j.synbio.2025.03.009","url":null,"abstract":"<div><div><em>In situ</em> imaging diagnosis and precise treatment of deep tumor tissues are hotspots in life sciences and medical research. In recent years, using focused ultrasound to remotely control engineered bacteria for drug release has become one of the methods for precise <em>in vivo</em> drug delivery. However, non-visualized engineered bacteria pose challenges for precise control within the body. Therefore, there is an urgent need for an engineered bacterial vector capable of deep tissue imaging to precisely locate bacteria <em>in vivo</em>. Acoustic reporter genes (ARGs) are biological elements used for deep tissue imaging, with gene clusters over 8 kb. However, ARGs are often tested on plasmids, which hinders stable expression <em>in vivo</em> and limits the space for inserting components that regulate drug release. Therefore, we used the attenuated <em>Salmonella typhimurium</em> VNP20009, known for its tumor-targeting ability, as the chassis bacteria. By using CRISPR-Cas9 technology, we inserted ARGs into the genome and optimized the promoter strength and copy number for ARG expression, constructing ultrasound-visible engineered bacteria expressing gas vesicles on the genome. Additionally, by knocking out the stress protein gene <em>htrA</em> in VNP20009, we increased the maximum injection dose by tenfold and the tumor specificity by a hundredfold. The constructed ultrasound-visible engineered bacteria can stably synthesize gas vesicles and output ultrasound signals while directly carrying drug plasmids for tumor therapy. Our research provides an effective vector for diagnosis and precise treatment.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 3","pages":"Pages 774-782"},"PeriodicalIF":4.4,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143816405","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}