Trends in biotechnology最新文献_第8页

Rapid selection of transgenic mammalian cells via diphtheria toxin resistance. 通过白喉毒素抗性快速选择转基因哺乳动物细胞。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-09-01 Epub Date: 2025-06-18 DOI: 10.1016/j.tibtech.2025.05.010

David Scherer, Steffen Honrath, Jean-Christophe Leroux, Michael Burger

{"title":"Rapid selection of transgenic mammalian cells via diphtheria toxin resistance.","authors":"David Scherer, Steffen Honrath, Jean-Christophe Leroux, Michael Burger","doi":"10.1016/j.tibtech.2025.05.010","DOIUrl":"10.1016/j.tibtech.2025.05.010","url":null,"abstract":"The ability to generate stable transgenic mammalian cell lines is crucial to the investigation of gene functions and the production of recombinant proteins. Mammalian cells can be readily transfected in cell culture settings via both viral and nonviral vectors to induce transgene expression. However, there is an unmet need for efficient selection of transfected cells, since current methods involve rather inefficient antibiotic selection protocols or require the coexpression of fluorescent marker proteins, followed by laborious cell-sorting procedures. Thus, our aim was to implement a rapid and efficient selection approach for transgene-expressing human cells, using an engineered diphtheria toxin (DT) resistance-based selection, referred to as selecDT. We demonstrated that selecDT is expressed on the cell surface, provides efficient protection from DT by inactivating its uptake receptor, and, therefore, enables selection. SelecDT allows for greater selection efficiency in a more rapid timeline compared with conventional antibiotic methods. Thus, the resistance described herein may positively impact biotechnological processes.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":"2362-2377"},"PeriodicalIF":14.9,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144326977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Highly efficient prime editors for mammalian genome editing based on porcine retrovirus reverse transcriptase. 基于猪逆转录病毒逆转录酶的高效哺乳动物基因组编辑器。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-30 DOI: 10.1016/j.tibtech.2025.07.029

Weiwei Liu, Wenxin Duan, Zhiwei Peng, Yaya Liao, Xiaoguo Wang, Ruirong Liu, Qiqi Jing, Haoyun Jiang, Yuhang Fan, Liming Ge, Lusheng Huang, Yuyun Xing

{"title":"Highly efficient prime editors for mammalian genome editing based on porcine retrovirus reverse transcriptase.","authors":"Weiwei Liu, Wenxin Duan, Zhiwei Peng, Yaya Liao, Xiaoguo Wang, Ruirong Liu, Qiqi Jing, Haoyun Jiang, Yuhang Fan, Liming Ge, Lusheng Huang, Yuyun Xing","doi":"10.1016/j.tibtech.2025.07.029","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.029","url":null,"abstract":"Prime editing is a versatile and precise genome-editing tool. Most prime editors (PEs) rely on reverse transcriptase (RT) derived from Moloney murine leukemia virus (MMLV). Here, we established a PE, pvPE, using a RT derived from a porcine endogenous retrovirus (PERV) from a Bama mini-pig. Through various optimization strategies, including RT engineering, structural modifications, and La protein fusion, we gradually upgraded to pvPE-V4. This version achieved 24.38-101.69-fold higher efficiency compared with pvPE-V1 and up to 2.39-fold higher efficiency than another upgraded PE, PE7, with significantly fewer unintended edits across multiple mammalian cell lines. We further show that nocodazole (Noc) significantly enhanced pvPE efficiency by 2.25-fold on average. Using our pvPE system, we efficiently modified three genes simultaneously in porcine fibroblasts and subsequently generated cloned pigs that could serve as valuable models for Alzheimer's disease (AD) in humans. Our results highlight the broad application prospects of pvPE systems in mammalian genome editing.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Engineering infrared light detection in blind human retina using ultrasensitive human TRPV1 channels. 利用超灵敏人体TRPV1通道对盲人视网膜进行工程红外光检测。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.tibtech.2025.07.033

Morgan Chevalier, Firas Fadel, Tímea Májer, Dániel Péter Magda, Lili Gerendás, Ferenc Kilin, Zoltán Zsolt Nagy, Arnold Szabó, Botond Roska, Guilherme Testa-Silva

{"title":"Engineering infrared light detection in blind human retina using ultrasensitive human TRPV1 channels.","authors":"Morgan Chevalier, Firas Fadel, Tímea Májer, Dániel Péter Magda, Lili Gerendás, Ferenc Kilin, Zoltán Zsolt Nagy, Arnold Szabó, Botond Roska, Guilherme Testa-Silva","doi":"10.1016/j.tibtech.2025.07.033","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.033","url":null,"abstract":"Engineering infrared light sensitivity in the blind human retina could restore visual function in patients with regional retinal degeneration. However, current approaches are complex and contain non-human biological components. Using rational protein design, we engineered human transient receptor potential vanilloid 1 (hTRPV1) channels (Δ786-840) with temperature sensitivity that shifted from 45 to 41°C, which enabled near-infrared (NIR) light-induced heat activation of mammalian cells at close to physiological temperatures. When expressed in ganglion cells of human retinal explants, Δ786-840 TRPV1 generated robust spiking responses to brief NIR light-induced temperature transients. In addition, increasing intensity of radiation evoked graded responses correlating with increasing firing frequencies. Unlike previous approaches, which used non-human TRPV1 channels, risking immune reactions, and a multicomponent system that poses barriers to clinical implementation, this single-component human-derived approach eliminates immunogenicity concerns, addressing a major challenge to clinical translation, and allows gene delivery using adeno-associated virus (AAV) vectors.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Targeting RAS-mutant cancer cells using a synthetic RAS-activated cancer killing system. 利用合成ras激活的癌症杀伤系统靶向ras突变癌细胞。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.tibtech.2025.07.031

Fei Teng, Qingqin Gao, Li Zhou, Tongtong Cui, Xiangtian Tan, Yali Ding, Rongqi Li, Bojin Li, Bei Zhong, Miao Miao, Qi Zhou, Wei Li

{"title":"Targeting RAS-mutant cancer cells using a synthetic RAS-activated cancer killing system.","authors":"Fei Teng, Qingqin Gao, Li Zhou, Tongtong Cui, Xiangtian Tan, Yali Ding, Rongqi Li, Bojin Li, Bei Zhong, Miao Miao, Qi Zhou, Wei Li","doi":"10.1016/j.tibtech.2025.07.031","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.031","url":null,"abstract":"Despite being the most commonly mutated proteins in cancer, oncogenic RAS proteins remain largely untapped as pharmacological targets. Here, we report a synthetic cancer-killing platform, termed 'RAS-activated cancer killing (RACK)' system. Leveraging a transcriptional sensor designed to detect oncogenic RAS signals with high specificity, RACK achieves targeted identification and elimination of RAS-mutant cancer cells. RACK can potently target a range of RAS and non-RAS mutants, including, but not limited to KRAS, NRAS, BRAF, and RTKs. Notably, RACK can maintain its efficacy against cancer cells that have developed acquired resistance, outperforming conventional inhibitors. In vivo, RACK selectively inhibits RAS-mutant tumor growth in xenograft models, including those intractable by allele-specific inhibitors. Furthermore, the modular design of RACK allows rational optimization of promoter inputs and therapeutic outputs. Collectively, RACK introduces a pioneering drug approach for detecting and treating RAS-mutant cancers, paving the way for overcoming challenges associated with currently undruggable cancer targets.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970741","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Extended human lymph node explants for evaluation of adaptive immunity. 用于评价适应性免疫的扩展人淋巴结外植体。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.tibtech.2025.07.020

Kanishka Fernando, Hong Sheng Quah, Lisda Suteja, Anne James, Fathima F Kuthubudeen, Kenny Z Wu, Christabella Adine, Hariraman Bhuvaneswari, Mohanaselvi Senthilkumar, Sathiyamoorthy Selvarajan, N Gopalakrishna Iyer, Eliza L S Fong

{"title":"Extended human lymph node explants for evaluation of adaptive immunity.","authors":"Kanishka Fernando, Hong Sheng Quah, Lisda Suteja, Anne James, Fathima F Kuthubudeen, Kenny Z Wu, Christabella Adine, Hariraman Bhuvaneswari, Mohanaselvi Senthilkumar, Sathiyamoorthy Selvarajan, N Gopalakrishna Iyer, Eliza L S Fong","doi":"10.1016/j.tibtech.2025.07.020","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.020","url":null,"abstract":"Lymph nodes (LNs) are a vital component of the adaptive immune system as they have a key role in antigen presentation and regulation of immune responses. However, preclinical models that accurately mimic the complexity and spatial organization of LNs remain a significant unmet need for the study of LN biology. Here, we leveraged the use of biomaterials to significantly extend the lifespan of patient-derived LN explants ex vivo. Hydrogel-embedded LN explants preserved the cellular composition and maintained the intricate spatial organization of the LN. This enabled the LN explants to retain functional responsiveness, as demonstrated by their ability to mount immune responses after exposure to tumor antigens or SARS-CoV-2 mRNA vaccine. The LN explant models developed in this study offer a robust and physiologically relevant platform for studying immune responses ex vivo, facilitating the development of vaccines and immunotherapies in the context of cancer and infectious diseases.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970704","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of a high-efficiency N-acetylneuraminic acid production platform through multi-pathway synergistic engineering. 利用多途径协同工程技术开发高效n -乙酰神经氨酸生产平台。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.tibtech.2025.07.016

Guozhen Ma, Xiaolong Jiang, Bin Yang, Linxing Li, Ruiying Liu, Qing Meng, Jiawei Li, Lijie Xie, Han Guo, Sijia Liu, Yuxuan Wang, Yibo Wang, Xiaojing Zhao, Ziyu Li, Yujie Wang, Menglei Xia, Di Huang

{"title":"Development of a high-efficiency N-acetylneuraminic acid production platform through multi-pathway synergistic engineering.","authors":"Guozhen Ma, Xiaolong Jiang, Bin Yang, Linxing Li, Ruiying Liu, Qing Meng, Jiawei Li, Lijie Xie, Han Guo, Sijia Liu, Yuxuan Wang, Yibo Wang, Xiaojing Zhao, Ziyu Li, Yujie Wang, Menglei Xia, Di Huang","doi":"10.1016/j.tibtech.2025.07.016","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.016","url":null,"abstract":"The growing demand for N-acetylneuraminic acid (NeuAc) has driven the need for efficient and environmentally sustainable biomanufacturing processes. Microbial fermentation offers a promising route, yet optimizing cell factories with excellent phenotypes remains challenging. Here, we engineered Escherichia coli to enable high-efficiency co-utilization of glucose and glycerol. We refactored two synthetic pathways with the same start and end to enhance N-acetylmannosamine (ManNAc) precursor levels and optimized NeuAc synthase using artificial intelligence (AI) techniques and machine learning (ML) sequence mining. Subsequently, phosphoenolpyruvate (PEP) levels were boosted by capturing carbon flow from competing regeneration pathways, thus balancing the intracellular PEP:ManNAc ratio for improved NeuAc synthesis. Besides glucose, an additional carbon inlet from glycerol was opened, achieving a NeuAc titer of 70.4 g/l in fed-batch fermentation with a productivity of 1.17 g/l/h. This work demonstrates a highly efficient microbial cell factory for the biosynthesis of NeuAc and provides a versatile system engineering strategy applicable to other high-value compounds.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Membrane-targeted DNA frameworks with biodegradability recover cellular function and morphology from frozen cells. 具有生物降解性的膜靶向DNA框架可从冷冻细胞中恢复细胞功能和形态。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.tibtech.2025.07.028

Yedam Lee, Woo Hyuk Jung, Kyounghwa Jeon, Eui Bum Choi, Taeyoung Ryu, Chanseok Lee, Do-Nyun Kim, Dong June Ahn

{"title":"Membrane-targeted DNA frameworks with biodegradability recover cellular function and morphology from frozen cells.","authors":"Yedam Lee, Woo Hyuk Jung, Kyounghwa Jeon, Eui Bum Choi, Taeyoung Ryu, Chanseok Lee, Do-Nyun Kim, Dong June Ahn","doi":"10.1016/j.tibtech.2025.07.028","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.028","url":null,"abstract":"Cell freezing is critical for the long-term preservation of biological materials, but is limited by the cytotoxicity and inefficacy of conventional cryoprotective agents, such as dimethyl sulfoxide (DMSO). Here, we introduce DNA frameworks (DFs) as a nanoengineered programmable class of cryoprotectants designed to address these challenges. The DFs feature a programmable scaffolded structure offering large flexible wireframe contacts, cellular target ability, and biodegradability. Cholesterol-functionalized DFs outperformed conventional cryoprotectants in the recovery and maintenance of cellular functionality and morphology of frozen cells. Their cryoprotective mechanism enables targeted binding to the cell membrane, minimizing intracellular penetration or uptake, inhibits intracellular and extracellular ice growths, and promotes efficient post-thaw degradation to mitigate toxicity risks. By combining membrane-targeting specificity, cryoprotective efficacy, and biocompatibility, these DFs represent a transformative advance in cell cryopreservation.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biomechanics in miniature: microfluidic-based in vitro modeling to decipher mechanobiological phenomena. 微型生物力学：基于微流体的体外建模来破译机械生物学现象。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.tibtech.2025.07.024

Hye Jin Hong, Na-Hyun Lee, Hye Sung Kim, Kam W Leong, Hae-Won Kim

引用次数: 0

Towards gene editing and torrefaction pretreatment for biochar functionalization. 生物炭功能化的基因编辑与预处理研究。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-28 DOI: 10.1016/j.tibtech.2025.07.030

Congyu Zhang, Kuifeng Hao, Wei-Hsin Chen, Jakub Čespiva, Jiří Ryšavý, Ying Zhang

引用次数: 0

Engineered RNA nanostructures for scalable and efficient RNAi-based pesticides. 用于可扩展和高效的RNA基农药的工程RNA纳米结构。

IF 14.9 1区工程技术

Trends in biotechnology Pub Date : 2025-08-25 DOI: 10.1016/j.tibtech.2025.07.027

Ying Xia, Tao Zhang, Xiaokun Ni, Ying Zhu, Qian Chen, Anfu Bamu, Huan Dai, Xinda Lin

{"title":"Engineered RNA nanostructures for scalable and efficient RNAi-based pesticides.","authors":"Ying Xia, Tao Zhang, Xiaokun Ni, Ying Zhu, Qian Chen, Anfu Bamu, Huan Dai, Xinda Lin","doi":"10.1016/j.tibtech.2025.07.027","DOIUrl":"https://doi.org/10.1016/j.tibtech.2025.07.027","url":null,"abstract":"Double-stranded RNA (dsRNA)-based pesticides face challenges in stability, scalability, efficient uptake, and broad applicability. Here, we present self-assembled RNA nanostructures (SARNs), engineered to load pools of functional siRNAs with motifs that enhance hydrophobicity and elasticity, and enable both immediate and sustained siRNA release for efficient RNAi. SARNs improve RNA stability and delivery in plants and in model pests with chewing mouthparts (Tribolium castaneum) and piercing-sucking mouthparts (Nilaparvata lugens). Compared with dsRNA, SARNs demonstrated superior RNAi efficiency in T. castaneum and N. lugens, achieving significantly higher downregulation efficacy and mortality in both species. In addition, SARNs, which self-assemble from single-stranded (ss)RNA molecules, can be transcribed in Escherichia coli for scalable production. We further establish a framework for the laboratory-to-field transition of SARNs. This engineered RNA platform offers an efficient, scalable, cost-effective solution for RNA-based gene silencing, advancing applications in agriculture and biomedicine.","PeriodicalId":23324,"journal":{"name":"Trends in biotechnology","volume":" ","pages":""},"PeriodicalIF":14.9,"publicationDate":"2025-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0