Trends in biotechnology最新文献

Promoters are DNA sequences that govern the location, direction, and strength of gene transcription, playing a pivotal role in cellular growth and lifespan. Engineered promoters facilitate precise control of recombinant protein expression and metabolic pathway modulation for natural product biosynthesis. Traditional methods such as rational design and directed evolution have established the foundation for promoter engineering, and recent advances in deep learning (DL) have revolutionized the field. This review highlights the application of DL techniques for promoter identification, strength prediction, and de novo design using generative models. We describe how these tools are used and the impact of database quality, feature extraction, and model architecture on predictive accuracy. We discuss challenges and perspectives in developing robust models for promoter engineering.

Diagnostic sensor technologies lie at the core of the healthcare system and encompass disease screening, detection, and therapy monitoring. In past decades, many nanobiosensor technologies have emerged which rely on diverse principles using electrical, magnetic, mass-based, or photonic signal transduction methods. We provide an overview of recent and emerging nanobiosensing transduction technologies and illustrate the reported quantification capabilities for nucleic acids, proteins, and small molecules. The review assesses and compares their performance, multimodality, and multiplexing capabilities as well as their portability and throughput, among other criteria. In addition, we elaborate on demonstrated as well as envisaged medical applications of nanobiosensors. Finally, fundamental limitations such as the diffusion limit are discussed and opportunities for future research are outlined.

Carbonic anhydrase (CA) enzymes hold strong potential in new biotechnological strategies for accelerated CO₂ capture and conversion. Some CAs naturally tolerate the harsh conditions associated with carbon capture technologies; however, long-term durability, while maintaining high activity, presents significant challenges. This review offers an in-depth analysis of the CA enzymes that have been investigated for industrial carbon capture processes and highlights the key amino acids and structural features that are crucial for CA activity and stability under harsh conditions. It examines the impact of site-directed protein engineering to enhance CA efficacy and immobilization strategies. Furthermore, it addresses the challenges of scaling up CA-based technologies and offers strategies to improve their functionality. Future research directions, including artificial intelligence (AI)-driven optimization, are also discussed.

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.

Biohybrid-engineered tissue (BHET) platforms represent a cutting-edge development in tissue engineering by integrating electronic components with engineered biological tissues to enable real-time monitoring, precise modulation, and enhancement of tissue functionality. Recently, biofabrication and biofunctional material advances have facilitated the design of BHET platforms that respond to intricate biological and bioelectrical signals to replicate sophisticated physiological processes. BHET platforms have potential in disease modeling, regenerative medicine, and personalized healthcare applications by bridging the gap between biological and electrical systems. In this review, we classify BHET platforms into three platforms (tissue-sensor, tissue-electromodulator, and tissue-communicator platforms) and explore future directions, including innovations in biofabrication and data-driven platform design alongside optimization to enhance the scalability and functionality of the engineered human tissues.

Extracellular vesicles (EVs) have gained significant attention as therapeutics, building from natural mechanisms of paracrine signaling. The field has evolved with substantial heterogeneity in methods to isolate and characterize EVs and new methods are needed to scale-up EV production for therapeutic use. In this study, we isolated EVs from four porcine donors of bone marrow-derived mesenchymal stromal cells (MSCs) via three different cell culture methods: standard tissue culture plates, a 3D printed perfusion bioreactor, and microcarriers in spinner flasks. We explored EV manufacturing yield, characteristics, and content via proteomics and RNAseq. The MSC donor and their cell culture method affected the yield of EVs produced, whereas the method of EV isolation dominated the clustering of protein and RNA contents. As a step towards therapeutic application, in vitro tubule formation and hypoxic cardiac spheroid contraction assays showed improvements in vasculogenesis and cardiac cell recovery, respectively, in the presence of EVs.

Chronic wounds present a significant clinical challenge due to their complex pathophysiology and resistance to standard treatments. A key obstacle in developing therapies is the lack of animal models that accurately mimic human chronic wound characteristics. Existing rodent models fail to replicate critical features, such as delayed re-epithelialization and unresolved inflammation, while larger animals, including porcine models, also fall short. Here, we introduce a novel glutaraldehyde-induced porcine model that mimics key aspects of human chronic wounds. Glutaraldehyde causes dermal toxicity, resulting in impaired structural integrity, oxidative stress, persistent inflammation, and bacterial colonization. Analyses showed features such as delayed healing, extracellular matrix (ECM) disruption, mitochondrial dysfunction, and chronic inflammatory responses. Comparative transcriptomic and lipidomic studies revealed shared signaling pathways and metabolite profiles with human venous leg and diabetic foot ulcers, highlighting the translational relevance of the model. This innovative platform offers valuable insights into chronic wound mechanisms and aids the development of effective targeted therapies.

The rise of complex biotherapeutics has introduced bottlenecks in production using mammalian cells. Clustered regularly interspaced short palindromic repeats (CRISPR)-based screens enable unbiased discovery of engineering targets that mitigate biomanufacturing-relevant constraints. This forum gives an overview of recent advances and remaining challenges in applying CRISPR screening to build robust, modality-specific cell factories.

While regulatory functions of mature miRNAs are well established, the functions of miRNAs* and their potential for genetic engineering in crop improvement remain underexplored. Here, we used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas9) to generate artificial miR2118a/b (amiR2118a/b) by editing miR2118a/b-5p and obtained several amir2118a/b mutants in soybean (Glycine max). miR2118a/b-5p modifications altered the secondary structure of precursor amiR2118a/b (pre-amiR2118a/b) and reduced mature miR2118a/b levels. These amir2118a/b mutants retained the ability to initiate biogenesis of phased small interfering RNAs (phasiRNAs), albeit with a reduced abundance compared with wild-type (WT) plants. Furthermore, these mutants upregulated the expression of genes related to growth and defense under normal and Pseudomonas syringae pv. glycinea (Psg)-infected conditions, respectively. Notably, two transgene-free amir2118 mutants exhibited enhanced resistance to Psg, soybean cyst nematode (SCN), and root-knot nematode (RKN), and achieved increased yield under pathogen-free field conditions. This study provides a strategy to generate artificial miRNAs (amiRNAs) for crop improvement through the CRISPR/Cas system by mutating miRNAs* in crops.

Gene therapy has emerged as a promising strategy for tissue regeneration, offering the potential to address the limitations of conventional treatments. In this review we present an overview of applications of gene therapy in tissue regeneration, emphasizing recent advancements and future directions. Our work addresses gaps in the current literature by examining developments in molecular biology and genetics, such as clustered regularly interspaced short palindromic repeats (CRISPR) gene editing, advances in 3D bioprinting, and progress in gene delivery for tissue engineering. We describe case studies and clinical trials that demonstrate the potential of gene therapy applications in tissue engineering. We conclude by highlighting challenges and future directions, including emerging technologies and personalized gene-based approaches for tissue engineering research.