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Life
Special Issues
Microorganisms Engineering and Gene-Editing Methods
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Microorganisms Engineering and Gene-Editing Methods

A special issue of Life (ISSN 2075-1729). This special issue belongs to the section "Genetics and Genomics".

Deadline for manuscript submissions: 30 August 2025 | Viewed by 624

Special Issue Editors

Dr. Jichao Wang

E-Mail Website
Guest Editor
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China
Interests: metabolic engineering; genome editing; bioinformatics; enzyme engineering
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Guang Zhao

E-Mail Website
Guest Editor
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China
Interests: metabolic engineering; biobased materials and chemicals; gene expression regulation; bacterial stress response
Special Issues, Collections and Topics in MDPI journals
Dr. Xia Zhao

E-Mail Website
Guest Editor
Sun Yat-sen University Cancer Center (SYSUCC) Guangzhou, China Army Medical University, Chongqing, China
Interests: microbiology; microbiome; bioinformatics

Special Issue Information

Dear Colleagues,

Engineered microorganisms have emerged as critical assets across diverse sectors, including industrial biotechnology, biomedicine, agriculture, and environmental science. The precise construction and optimization of these strains necessitate advanced gene-editing techniques. Gene editing is not only fundamental to scientific progress but has also dramatically transformed our understanding of microorganisms. While significant progress has been achieved in recent decades, there remains considerable potential for refining these techniques to expand their applicability and address unmet challenges in various fields.

This Special Issue, titled "Microorganisms Engineering and Gene-Editing Methods", aims to present a comprehensive collection of articles exploring various facets of microorganism gene editing and its applications. The topics covered in this Issue include, but are not limited to the following:

  • Novel gene-editing techniques for bacteria, archaea, fungi, and other microbial systems.
  • Improvements in editing efficiency, specificity, and scalability, including strategies to minimize off-target effects and cytotoxicity.
  • Advancements in workflow optimization, such as transformation protocols, recombination systems, counter-selection methods, recombinase engineering, and plasmid curing.
  • Emerging CRISPR-based technologies, including CRISPR interference (CRISPRi), CRISPR activation (CRISPRa), base editors, prime editors, RNA-targeting systems, and CRISPR-associated transposase tools.
  • Metabolic engineering, strain optimization for bioproduction, functional analysis of microbial pathways, and the enhancement of traits through gene editing. The precise expression of exogenous genes to endow microorganisms with new characteristics is also encouraged.

We encourage researchers from academia and industry to contribute their work, fostering interdisciplinary dialogue and accelerating the translation of microbial engineering into real-world solutions.

Dr. Jichao Wang
Prof. Dr. Guang Zhao
Dr. Xia Zhao
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Life is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • microorganisms engineering
  • metabolic engineering
  • strain optimization
  • gene editing
  • CRISPR
  • recombination

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Published Papers (1 paper)

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Research

18 pages, 2158 KiB  
Article
Biosynthesis of Two Types of Exogenous Antigenic Polysaccharides in a Single Escherichia coli Chassis Cell
by Jingjing Hao, Haoqian Liao, Shuhong Meng, Yan Guo, Li Zhu, Hengliang Wang and Yufei Lyu
Life 2025, 15(6), 858; https://doi.org/10.3390/life15060858 - 26 May 2025
Viewed by 335
Abstract
Escherichia coli and Klebsiella pneumoniae are major contributors to the global challenge of antimicrobial resistance, posing serious threats to public health. Among current preventive strategies, conjugate vaccines that utilize bacterial surface polysaccharides have emerged as a promising and effective approach to counter multidrug-resistant [...] Read more.
Escherichia coli and Klebsiella pneumoniae are major contributors to the global challenge of antimicrobial resistance, posing serious threats to public health. Among current preventive strategies, conjugate vaccines that utilize bacterial surface polysaccharides have emerged as a promising and effective approach to counter multidrug-resistant strains. In this study, both the Wzy/Wzx-dependent and ABC transporter-dependent biosynthetic pathways for antigenic polysaccharides were introduced into E. coli W3110 cells. This dual-pathway engineering enabled the simultaneous biosynthesis of two structurally distinct polysaccharides within a single host, offering a streamlined and potentially scalable strategy for vaccine development. Experimental findings confirmed that both polysaccharide types were successfully produced in the engineered strains, although co-expression levels were moderately reduced. A weak competitive interaction was noted during the initial phase of induction, which may be attributed to competition for membrane space or the shared use of activated monosaccharide precursors. Interestingly, despite a reduction in plasmid copy number and transcriptional activity of the biosynthetic gene clusters over time, the overall polysaccharide yield remained stable with prolonged induction. This suggests that extended induction does not adversely affect final product output. Additionally, two glycoproteins were efficiently generated through in vivo bioconjugation of the synthesized polysaccharides with carrier proteins, all within the same cellular environment. This one-cell production system simplifies the workflow and enhances the feasibility of generating complex glycoprotein vaccines. Whole-cell proteomic profiling followed by MFUZZ clustering and Gene Ontology analysis revealed that core biosynthetic genes were grouped into two functional clusters. These genes were predominantly localized to the cytoplasm and were enriched in pathways related to translation and protein binding. Such insights not only validate the engineered biosynthetic routes but also provide a molecular basis for optimizing future constructs. Collectively, this study presents a robust synthetic biology platform for the co-expression of multiple polysaccharides in a single bacterial host. The approach holds significant promise for the rational design and production of multivalent conjugate vaccines targeting drug-resistant pathogens. Full article
(This article belongs to the Special Issue Microorganisms Engineering and Gene-Editing Methods)
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