Genetic Engineering Research Articles

Cadmium (Cd) Tolerance and Phytoremediation Potential in Fiber Crops: Research Updates and Future Breeding Efforts

Heavy metal pollution is one of the most devastating abiotic factors, significantly damaging crops and human health. One of the serious problems it causes is a rise in cadmium (Cd) toxicity. Cd is a highly toxic metal with a negative biological role, and it enters plants via the soil–plant system. Cd stress induces a series of disorders in plants’ morphological, physiological, and biochemical processes and initiates the inhibition of seed germination, ultimately resulting in reduced growth. Fiber crops such as kenaf, jute, hemp, cotton, and flax have high industrial importance and often face the issue of Cd toxicity. Various techniques have been introduced to counter the rising threats of Cd toxicity, including reducing Cd content in the soil, mitigating the effects of Cd stress, and genetic improvements in plant tolerance against this stress. For decades, plant breeders have been trying to develop Cd-tolerant fiber crops through the identification and transformation of novel genes. Still, the complex mechanism of Cd tolerance has hindered the progress of genetic breeding. These crops are ideal candidates for the phytoremediation of heavy metals in contaminated soils. Hence, increased Cd uptake, accumulation, and translocation in below-ground parts (roots) and above-ground parts (shoots, leaves, and stems) can help clean agricultural lands for safe use for food crops. Earlier studies indicated that reducing Cd uptake, detoxification, reducing the effects of Cd stress, and developing plant tolerance to these stresses through the identification of novel genes are fruitful approaches. This review aims to highlight the role of some conventional and molecular techniques in reducing the threats of Cd stress in some key fiber crops. Molecular techniques mainly involve QTL mapping and GWAS. However, more focus has been given to the use of transcriptome and TFs analysis to explore the potential genomic regions involved in Cd tolerance in these crops. This review will serve as a source of valuable genetic information on key fiber crops, allowing for further in-depth analyses of Cd tolerance to identify the critical genes for molecular breeding, like genetic engineering and CRISPR/Cas9.

Can nanotechnology and genomics innovations trigger agricultural revolution and sustainable development?

At the dawn of new millennium, policy makers and researchers focused on sustainable agricultural growth, aiming for food security and enhanced food quality. Several emerging scientific innovations hold the promise to meet the future challenges. Nanotechnology presents a promising avenue to tackle the diverse challenges in agriculture. By leveraging nanomaterials, including nano fertilizers, pesticides, and sensors, it provides targeted delivery methods, enhancing efficacy in both crop production and protection. This integration of nanotechnology with agriculture introduces innovations like disease diagnostics, improved nutrient uptake in plants, and advanced delivery systems for agrochemicals. These precision-based approaches not only optimize resource utilization but also reduce environmental impact, aligning well with sustainability objectives. Concurrently, genetic innovations, including genome editing and advanced breeding techniques, enable the development of crops with improved yield, resilience, and nutritional content. The emergence of precision gene-editing technologies, exemplified by CRISPR/Cas9, can transform the realm of genetic modification and enabled precise manipulation of plant genomes while avoiding the incorporation of external DNAs. Integration of nanotechnology and genetic innovations in agriculture presents a transformative approach. Leveraging nanoparticles for targeted genetic modifications, nanosensors for early plant health monitoring, and precision nanomaterials for controlled delivery of inputs offers a sustainable pathway towards enhanced crop productivity, resource efficiency, and food safety throughout the agricultural lifecycle. This comprehensive review outlines the pivotal role of nanotechnology in precision agriculture, emphasizing soil health improvement, stress resilience against biotic and abiotic factors, environmental sustainability, and genetic engineering.

The potent PHL4 transcription factor effector domain contains significant disorder.

The phosphate-starvation response transcription-factor protein family is essential to plant response to low-levels of phosphate. Proteins in this transcription factor (TF) family act by altering various gene expression levels, such as increasing levels of the acid phosphatase proteins which catalyze the conversion of inorganic phosphates to bio-available compounds. There are few structural characterizations of proteins in this TF family, none of which address the potent TF activation domains. The phosphate-starvation response-like protein-4 (PHL4) protein from this family has garnered interest due to the unusually high TF activation activity of the N-terminal domain. Here, we demonstrate using solution nuclear magnetic resonance (NMR) measurements that the PHL4 N-terminal activating TF effector domain is mainly an intrinsically disordered domain of over 200 residues, and that the C-terminal region of PHL4 is also disordered. Additionally, we present evidence from size-exclusion chromatography, diffusion NMR measurements, and a cross-linking assay suggesting full-length PHL4 forms a trimeric or tetrameric assembly. Together, the data indicate the N- and C-terminal disordered domains in PHL4 flank a central folded region that likely forms the ordered oligomer of PHL4. This work provides a foundation for future studies detailing how the conformations and molecular motions of PHL4 change as it acts as a potent activator of gene expression in phosphate metabolism. Such a detailed mechanistic understanding of TF function will benefit genetic engineering efforts that take advantage of this activity to boost transcriptional activation of genes across different organisms.

The Impact of Biotechnology in Shaping the Modern Agriculture

Advances in biotechnology have created new opportunities for food diversity in terms of both quantity and quality, as well as for collecting food in proportion to the world's expanding population and ensuring food security. With the advancement of molecular biology, DNA sequencing, and recombinant DNA in the middle of the 20th century, modern-day advancement of molecular breeding gave rise to genetic engineering and recombinent DNA techniques that are today frequently connected to biotechnology. This expansion has been facilitated by a variety of fields, including genetics, molecular biology, and biotechnology. The use of genetic engineering in agriculture has grown due to its ability to precisely create and insert a gene from any living organism. Molecular markers are also utilized to identify disease and pest resistance in order to increase agricultural output using traditional techniques. Genetic engineering plays a crucial role in agriculture, representing the scientific and technical advancement of traditional methods for crop enhancement. Recombinant DNA technology, RNA interference, and CRISPR genomic editing are some of the instruments and methods used in genetic engineering. The emergence of biotechnological innovations aimed at improving the nutritional composition of individual foods has led to the growing problem of a malnourished world population, where a growing number of people rely on large regions of the world for food used as the only source of protein, carbohydrates, fats and several essential minerals and vitamins, in addition to being a source of energy.

Revolutionizing dairy waste: emerging solutions in conjunction with microbial engineering.

The dairy industry is grappling with significant challenges in managing effluent due to environmental concerns and stringent regulatory demands, necessitating innovative solutions. The paper investigates how microbial engineering is transforming the treatment of dairy wastewater, offering advanced methods to minimize environmental impact and enhance sustainability. It delves into the current challenges faced by the dairy industry, such as regulatory compliance and the limitations of traditional treatment technologies, and introduces microbial engineering as a promising solution for effluent management. Microbial engineering leverages genetic engineering techniques and microorganisms to enhance the efficiency of treatment processes like bioaugmentation and bioremediation. The environmental and economic benefits of microbial engineering, highlighting its potential to reduce pollution and lower operational costs for the dairy industry. The specific figures can vary based on factors like farm size and location, studies suggest that microbial engineering can reduce wastewater pollution by up to 50% and nutrient runoff by 30%. It also identifies key challenges and there are still areas including strains for specific pollutants (drugs, hormones), enhance degradation pathways, and increase microbes' stability (stress tolerance, long-term viability) that require further innovation to maximize its benefits. Through case studies and success stories, the paper demonstrates practical applications of microbial engineering in managing dairy effluent, illustrating how it can revolutionize industrial practices for a more sustainable future.

Genetic Engineering in Bacteria, Fungi, and Oomycetes, Taking Advantage of CRISPR

Genetic engineering has revolutionized our ability to modify microorganisms for various applications in agriculture, medicine, and industry. This review examines recent advances in genetic engineering techniques for bacteria, fungi, and oomycetes, with a focus on CRISPR-Cas systems. In bacteria, CRISPR-Cas9 has enabled precise genome editing, enhancing applications in antibiotic production and metabolic engineering. For fungi, despite challenges associated with their complex cell structures, CRISPR/Cas9 has advanced the production of enzymes and secondary metabolites. In oomycetes, significant plant pathogens, modified Agrobacterium-mediated transformation, and CRISPR/Cas12a have contributed to developing disease-resistant crops. This review provides a comparative analysis of genetic engineering efficiencies across these microorganisms and addresses ethical and regulatory considerations. Future research directions include refining genetic tools to improve efficiency and expand applicability in non-model organisms. This comprehensive overview highlights the transformative potential of genetic engineering in microbiology and its implications for addressing global challenges in agriculture, medicine, and biotechnology.

Construction of engineered probiotic that adhere and display nanobody to neutralize porcine reproductive and respiratory syndrome virus.

Pathogenic blue ear disease caused by porcine reproductive and respiratory syndrome virus (PRRSV) bring severe loss to breeding industry due to high infectivity and mortality. L. plantarum serves as the probiotic host strain, known for its beneficial properties in the gut microbiota. E. coli is used as a cloning host for the initial genetic engineering steps, facilitating the construction and amplification of the desired genetic constructs. In this study, using synthetic biology technology, we constructed engineered probiotics which could adhere and display nanobody on the surface to neutralize virus. Firstly, we screen an optimal nanobody to effectively bind with PRRSV by building library, expression and purification. Then, the integration of adhesion protein and nanobody into the genome of probiotics significantly improved its adhesion to IPEC-J2 cells. In addition, this engineered probiotic is almost non-toxic to cells with good safety, which can be used as a daily probiotics to prevent virus fecal transmission. Our study proposed this novel construction strategy of engineering probiotics with both adhesion and neutralization effects, which provided a new therapeutic view for intestinal virus clearance.

Engineering Saccharomyces cerevisiae for continuous secretory production of hEGF in biofilm.

Human epidermal growth factor (hEGF) plays a crucial role in promoting cell growth and has various clinical applications. Due to limited natural sources and the high cost of chemical synthesis, researchers are now exploring genetic engineering as a potential method for hEGF production. In this particular study, a novel hEGF expression system was developed using Saccharomyces cerevisiae. This system involved optimizing the promoter and signal peptide and deleting protease-coding genes PEP4, PRB1, and YAP3, overexpressing chaperones KAR2 and PDI1 in the protein secretion pathway, which led to a 2.01-fold increase in hEGF production compared to the wild type strain. Furthermore, biofilm-forming genes FLO11 and ALS3 were integrated to create a biofilm strain with adhesive properties. A biofilm-based immobilized continuous fermentation model was established to leverage the characteristics of this biofilm strain. Each batch of this model yielded 130mg/L of hEGF, with a production efficiency of 2.71mg/L/h - surpassing the production efficiency of traditional free fermentation (1.62mg/L/h). This study presents a promising fermentation model for efficient hEGF production based on biofilm characteristics, offering valuable insights for the application of biofilm fermentation in the production of small molecule peptides.

Abstract A012: g3bp1-mediated stress granule formation drives melanoma initiation in zebrafish

Abstract Stress granules confer cancer cells the ability to withstand harsh biological conditions and impart tumorigenic translational states through mRNA sequestration. Despite advances in in vitro knowledge, mechanistic understanding of the endogenous influence of stress granules during tumorigenesis in vivo remains understudied. To address this, we use a zebrafish melanoma model where oncogenic human BRAF V600E is driven by the melanocyte master regulator mitfa in a p53 -/- background. Through vector-based genetic engineering, endogenous melanoma induction in these zebrafish can be developmentally staged from single cell to tumor. BRAF V600E ::p53 -/- melanocytes form a cancerized field (CF), from which a subset upregulate the melanocyte master regulator mitfa, creating a cancer precursor zone (CPZ). Transformation of CPZ cells to an embryonic, neural crest-like state facilitates their formation of an early tumor patch that inevitable develops into an overt tumor. Single cell RNAseq analysis of cells representing each melanoma stage showed that CPZ cells upregulate core stress granule markers, including g3bp1, tia1, tia1l, and nufip2. Immunostaining of g3bp1 in CF, CPZ, patch, and tumor cells revealed a significant induction of stress granules from the CF to CPZ transition (2 vs 10 stress granules/cell; N=3; p=0.034) that persists during the patch and tumor stage, indicating cell stress accompanies melanoma initiation and progression. Genetic disruption of g3bp1 through cell autonomous, melanocyte-specific CRISPR editing in zebrafish (N=10-13) causes delayed CPZ onset (P=0.012) and fewer CPZs to form (P=0.021). In turn, g3bp1-edited zebrafish have delayed tumor formation (P=0.026) and develop fewer tumors (P=0.006). To decipher the RNAs sequestered by stress granules during melanomagenesis, we performed RIPseq for G3BP1 in human A375 melanoma cells stressed with sodium arsenite. RNAs significantly bound to G3BP1 upon stress induction include major tumor suppressors, such as CDKN2A, RBM5, BIK, and RHOB. Together, these results suggest that endogenous g3bp1-mediated stress granule formation in melanoma initiating cells is tumorigenic in vivo and operates through the sequestration of tumor suppressive RNAs. Citation Format: Kyle D Drake, Emily Formato, Leonard Zon. g3bp1-mediated stress granule formation drives melanoma initiation in zebrafish [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr A012.

Significant Advancements and Evolutions in Chimeric Antigen Receptor Design

Recent times have witnessed remarkable progress in cancer immunotherapy, drastically changing the cancer treatment landscape. Among the various immunotherapeutic approaches, adoptive cell therapy (ACT), particularly chimeric antigen receptor (CAR) T cell therapy, has emerged as a promising strategy to tackle cancer. CAR-T cells are genetically engineered T cells with synthetic receptors capable of recognising and targeting tumour-specific or tumour-associated antigens. By leveraging the intrinsic cytotoxicity of T cells and enhancing their tumour-targeting specificity, CAR-T cell therapy holds immense potential in achieving long-term remission for cancer patients. However, challenges such as antigen escape and cytokine release syndrome underscore the need for the continued optimisation and refinement of CAR-T cell therapy. Here, we report on the challenges of CAR-T cell therapies and on the efforts focused on innovative CAR design, on diverse therapeutic strategies, and on future directions for this emerging and fast-growing field. The review highlights the significant advances and changes in CAR-T cell therapy, focusing on the design and function of CAR constructs, systematically categorising the different CARs based on their structures and concepts to guide researchers interested in ACT through an ever-changing and complex scenario. UNIVERSAL CARs, engineered to recognise multiple tumour antigens simultaneously, DUAL CARs, and SUPRA CARs are some of the most advanced instances. Non-molecular variant categories including CARs capable of secreting enzymes, such as catalase to reduce oxidative stress in situ, and heparanase to promote infiltration by degrading the extracellular matrix, are also explained. Additionally, we report on CARs influenced or activated by external stimuli like light, heat, oxygen, or nanomaterials. Those strategies and improved CAR constructs in combination with further genetic engineering through CRISPR/Cas9- and TALEN-based approaches for genome editing will pave the way for successful clinical applications that today are just starting to scratch the surface. The frontier lies in bringing those approaches into clinical assessment, aiming for more regulated, safer, and effective CAR-T therapies for cancer patients.

Precision Agriculture and Water Conservation Strategies for Sustainable Crop Production in Arid Regions

The intensifying challenges posed by global climate change and water scarcity necessitate enhancements in agricultural productivity and sustainability within arid regions. This review synthesizes recent advancements in genetic engineering, molecular breeding, precision agriculture, and innovative water management techniques aimed at improving crop drought resistance, soil health, and overall agricultural efficiency. By examining cutting-edge methodologies, such as CRISPR/Cas9 gene editing, marker-assisted selection (MAS), and omics technologies, we highlight efforts to manipulate drought-responsive genes and consolidate favorable agronomic traits through interdisciplinary innovations. Furthermore, we explore the potential of precision farming technologies, including the Internet of Things (IoT), remote sensing, and smart irrigation systems, to optimize water utilization and facilitate real-time environmental monitoring. The integration of genetic, biotechnological, and agronomic approaches demonstrates a significant potential to enhance crop resilience against abiotic and biotic stressors while improving resource efficiency. Additionally, advanced irrigation systems, along with soil conservation techniques, show promise for maximizing water efficiency and sustaining soil fertility under saline–alkali conditions. This review concludes with recommendations for a further multidisciplinary exploration of genomics, sustainable water management practices, and precision agriculture to ensure long-term food security and sustainable agricultural development in water-limited environments. By providing a comprehensive framework for addressing agricultural challenges in arid regions, we emphasize the urgent need for continued innovation in response to escalating global environmental pressures.

Gene expression screening and cell factory engineering for enhancing echinocandin B production in Aspergillus nidulans NRRL8112.

Echinocandin B (ECB) is a key precursor of the antifungal drug anidulafungin and its biosynthesis occurs via ani gene cluster in Aspergillus nidulans NRRL8112. Strain improvement for industrial ECB production has mainly relied on mutation breeding due to the lack of genetic tools. Here, a CRISPR-base-editing tool was developed in A. nidulans NRRL8112 for simultaneous inactivation of the nkuA gene and two marker genes, pryoA and riboB, which enabled efficient genetic manipulation. Then, in-vivo plasmid assembly was harnessed for ani gene expression screening, identifying the rate-limiting enzyme AniA and a pathway-specific transcription factor AniJ. Stepwise titer enhancement was achieved by overexpressing aniA and/or aniJ, and ECB production reached 1.5g/L during 5-L fed-batch fermentation, an increase of ~ 30-fold compared with the parent strain. This study, for the first time, revealed the regulatory mechanism of ECB biosynthesis and harnessed genetic engineering for the development of an efficient ECB-producing strain.

Creeping Stem 1 regulates directional auxin transport for lodging resistance in soybean.

Soybean, a staple crop on a global scale, frequently encounters challenges due to lodging under high planting densities, which results in significant yield losses. Despite extensive research, the fundamental genetic mechanisms governing lodging resistance in soybeans remain elusive. In this study, we identify and characterize the Creeping Stem 1 (CS1) gene, which plays a crucial role in conferring lodging resistance in soybeans. The CS1 gene encodes a HEAT-repeat protein that modulates hypocotyl gravitropism by regulating amyloplast sedimentation. Functional analysis reveals that the loss of CS1 activity disrupts polar auxin transport, vascular bundle development and the biosynthesis of cellulose and lignin, ultimately leading to premature lodging and aberrant root development. Conversely, increasing CS1 expression significantly enhances lodging resistance and improves yield under conditions of high planting density. Our findings shed light on the genetic mechanisms that underlie lodging resistance in soybeans and highlight the potential of CS1 as a valuable target for genetic engineering to improve crop lodging resistance and yield.

Development of an Escherichia coli Cell-Based Biosensor for Aspirin Monitoring by Genetic Engineering of MarR

Multiple antibiotic resistance regulators (MarRs) control the transcription of genes in the mar operon of Escherichia coli in the presence of salicylic acid (SA). The interaction with SA induces conformational changes in the MarR released from the promoter of the mar operon, turning on transcription. We constructed an SA-specific E. coli cell-based biosensor by fusing the promoter of the mar operon (PmarO) and the gene that encodes an enhanced green fluorescent protein (egfp). Because SA and aspirin are structurally similar, a biosensor for monitoring aspirin can be obtained by genetically engineering MarR to be aspirin (ASP)-responsive. To shift the selectivity of MarR toward ASP, we changed the residues around the ligand-binding sites by site-directed mutagenesis. We examined the effects of genetic engineering on MarR by introducing MarRs with PmarO-egfp into E. coli. Among the tested mutants, MarR T72A improved the ASP responses by approximately 3 times compared to the wild-type MarR, while still showing an SA response. Although the MarR T72A biosensor exhibited mutual interference between SA and ASP, it accurately determined the ASP concentration in spiked water and medicine samples with over 90% accuracy. While the ASP biosensors still require improvement, our results provide valuable insights for developing E. coli cell-based biosensors for ASP and transcription factor-based biosensors in general.

The Role of Biotechnology in Shaping the Future of Modern Agriculture

Biotechnology stands as a cornerstone in the evolution of modern agriculture, offering an array of innovative solutions poised to revolutionize crop productivity, enhance nutritional quality, and foster environmental sustainability. This comprehensive review delves into the multifaceted role of biotechnology in shaping the future landscape of agriculture, encompassing diverse realms such as genetic engineering, molecular breeding, and microbial biotechnology. Spanning from crop improvement to pest and disease management, soil health, and sustainable agriculture practices, the applications of biotechnology in agriculture are far-reaching and profound. Genetic engineering emerges as a powerful tool in the arsenal of agricultural biotechnology, facilitating the precise manipulation of plant genomes to confer desirable traits such as increased yield, enhanced nutritional content, and resistance to biotic and abiotic stresses. Molecular breeding techniques complement this approach, leveraging advances in genomics and bioinformatics to expedite the breeding process and develop high-performing crop varieties tailored to specific agroecological contexts, microbial biotechnology emerges as a pivotal force in sustainable agriculture, harnessing the power of beneficial microorganisms to improve soil fertility, enhance nutrient uptake, and suppress plant pathogens. From biofertilizers and biopesticides to bioremediation and phytoremediation, microbial-based solutions hold immense potential in mitigating environmental degradation and promoting agroecosystem resilience, the widespread adoption of biotechnology in agriculture is not devoid of societal, economic, and ethical considerations. Regarding food safety, environmental impact, and socio-economic equity underscore the need for rigorous regulatory frameworks and robust risk assessment protocols to ensure the responsible deployment of biotechnological innovations, public perceptions and fostering dialogue between stakeholders are essential for building trust and garnering support for biotechnology in agriculture. Education, transparency, and stakeholder engagement are paramount in navigating the complex terrain of biotechnology governance and fostering an enabling environment for innovation, the current state and future prospects of biotechnology in agriculture, this review endeavors to inform policymakers, researchers, and stakeholders about the transformative potential of biotechnology to address pressing global challenges such as food security, climate change, and environmental sustainability. Embracing the opportunities afforded by biotechnology holds the key to unlocking a more resilient, equitable, and sustainable agricultural future.

Non-replicative herpes simplex virus genomic and amplicon vectors for gene therapy - an update.

Two major types of defective vectors have been derived from herpes simplex virus type 1 (HSV-1), non-replicative genomic vectors (nrHSV-1), and amplicon vectors. This review recapitulates the main features of both vector types and summarizes recent improvements in our understanding of virus/vector biology, particularly with regard to the critical role played by the overpowering of antiviral cellular defenses and the epigenetic control of viral gene expression. Over the past years, significant breakthroughs in vector design, genetic engineering, and HSV-1 biology have accelerated the development of nrHSV-1 vectors. The low immunogenicity and enhanced safety profiles allowed the successful translation of these vectors into several clinical trials, with some being approved by the FDA. Regarding amplicons, despite their advantage in carrying very large or multiple transgenes, and their potential to avoid genome dilution in dividing cells, the absence of production procedures capable of generating large amounts of helper-free amplicons at reasonable cost with GMP compliance, still limits the translation of these outstanding vectors to clinical trials.

Genome-wide analysis of miR172-mediated response to heavy metal stress in chickpea (Cicer arietinum L.): physiological, biochemical, and molecular insights.

Chickpea (Cicer arietinum L.), a critical diploid legume in the Fabaceae family, is a rich source of protein, vitamins, and minerals. However, heavy metal toxicity severely affects its growth, yield, and quality. MicroRNAs (miRNAs) play a crucial role in regulating plant responses to both abiotic and biotic stress, including heavy metal exposure, by suppressing the expression of target genes. Plants respond to heavy metal stress through miRNA-mediated regulatory mechanisms at multiple physiological, biochemical, and molecular levels. Although the Fabaceae family is well represented in miRNA studies, chickpeas have been notably underrepresented. This study aimed to investigate the effects of heavy metal-induced stress, particularly from 100 µM concentrations of cadmium (Cd), chromium (Cr), nickel (Ni), lead (Pb), and 30 µM arsenic (As), on two chickpea varieties: ILC 482 (sensitive) and Azkan (tolerant). The assessment focused on physiological, biochemical, and molecular parameters. Furthermore, a systematic characterization of the miR172 gene family in the chickpea genome was conducted to better understand the plant's molecular response to heavy metal stress. Variance analysis indicated significant effects of genotype (G), treatment (T), and genotype-by-treatment (GxT) interactions on plant growth, physiological, and biochemical parameters. Heavy metal stress negatively impacted plant growth in chickpea genotypes ILC 482 and Azkan. A reduction in chlorophyll content and relative leaf water content was observed, along with increased cell membrane damage. In ILC 482, the highest hydrogen peroxide (H₂O₂) levels in shoot tissue were recorded under As, Cd, and Ni treatments, while in Azkan, peak levels were observed with Pb treatment. Malondialdehyde (MDA) levels in root tissue were highest in ILC 482 under Cd and Ni exposure and in Azkan under As, Cr, and Cd treatments. Antioxidant enzyme activities, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX), were significantly elevated under heavy metal stress in both genotypes. Gene expression analysis revealed upregulation of essential antioxidant enzyme genes, such as SOD, CAT, and APX, with APX showing notable increases in both shoot and root tissues compared to the control. Additionally, seven miR172 genes (miR172a, miR172b, miR172c, miR172d, miR172e, miR172f, and miR172g) were identified in the chickpea genome, distributed across five chromosomes. All genes exhibited conserved hairpin structures essential for miRNA functionality. Phylogenetic analysis grouped these miR172 genes into three clades, suggesting strong evolutionary conservation with other plant species. The expression analysis of miR172 and its target genes under heavy metal stress showed varied expression patterns, indicating their role in enhancing heavy metal tolerance in chickpea. Heavy metal stress significantly impaired plant growth and physiological and biochemical parameters in chickpea genotypes, except for cell membrane damage. The findings underscore the critical role of miR172 and its target genes in modulating chickpea's response to heavy metal stress. These insights provide a foundational understanding for developing stress-tolerant chickpea varieties through miRNA-based genetic engineering approaches.

A Brief Chronicle of Antibody Research and Technological Advances

This review briefly traces the historical development of antibody research and related technologies. The path from early perceptions of immunity to the emergence of modern immunotherapy has been marked by pivotal discoveries and technological advances. Early insights into immunity led to the development of vaccination and serotherapy. The elucidation of antibody structure and function paved the way for monoclonal antibody technology and its application in diagnosis and therapy. Breakthroughs in genetic engineering have enabled the production of humanized antibodies and the advances in Fc engineering, thereby increasing therapeutic efficacy. The discovery of immune checkpoints and cytokines revolutionized the treatment of cancer and autoimmune diseases. The field continues to evolve rapidly with the advent of antibody–drug conjugates, bispecific antibodies, and CAR T-cell therapies. As we face global health challenges, antibody research remains at the forefront of medical innovation and offers promising solutions for the future.

Antibacterial Activity of Silver Nanoparticles Prepared from Camellia Sinensis Extracts in Multi-Drug Resistant Staphylococcus Aureus.

The nano-method has been used as a technique for creating novel, non-traditional antimicrobial agents. This effective method for treating infectious diseases has many advantages over conventional antibiotics, including increased efficacy against species that have developed drug resistance, and the ability to circumvent the development of resistance that disrupts a number of biological pathways. As a result, the objective of this study was to synthesize and characterize silver nanoparticles using phenolic compounds obtained from Camellia sinensis . The nanoparticles were then used as antibacterial agents on the multidrug resistant Staphylococcus aureus, as well as, biofilm formation mechanism were also investigated. Ten isolates of Staphylococcus aureus were acquired from the labs of the University of Baghdad's Genetic Engineering and Biotechnology Institute. The VITEK-2 system was used to confirm the diagnosis. Aqueous and methanolic extracts of Camellia sinensis leaves were used to create silver nanoparticles and obtain CAgNPs, which were then characterized using Atomic Fluorescence Microscopy (AFM), X-ray diffractometer (XRD), and Zeta potential analyzers. The extracts were put through a series of assays, including High-performance liquid chromatography (HPLC), antibacterial activity assessments, and the microtiter plate method to determine the lowest inhibitory concentration (MIC) and antibiofilm formation. Both aqueous and methanolic extracts containing silver nanoparticles included spherical nanoparticles that may be single or combined. The HPLC results showed the presence of two phenolic compounds (gallic acid and caffeine) by comparing their retention durations to those of the reference compounds. The results of the antibacterial activity of (CAgNPs) showed that the methanolic (CAgNPs) extract was more effective than the aqueous (CAgNPs) extract, producing inhibitory zones of 15.67 ± 0.58 and 20.33 ± 0.58 mm, at 375 and 750 ppm respectively, when compared to the aqueous (CAgNPs) extract, which produced inhibitory zones of 12.33 ± 0.58 and 15.67 ± 0.58 mm, respectively. The MIC result also showed that the CAgNPs methanolic extract was more effective than the CAgNPs aqueous extract. The MIC of the CAgNPs methanolic extract on S. aureus isolates were 11.718 and 23.43 µg/ml, while the MIC of the CAgNPs aqueous extract on all S. aureus isolates were 46.87 µg/ml except isolate No. 3 and 6 which was 11.718 and 93.75 µg/ml respectively. Additionally, The anti-biofilm in S. aureus was increased when CAgNPs methanolic extract were used compared with the CAgNPs aqueous extract, the CAgNPs methanolic extract inhibited 80%, 90% and 100% of the biofilm formation of S. aureus in 23.43, 46.87 and 93.75 µg/ml respectively, while the anti-biofilm activity of the CAgNPs aqueous extract on S. aureus isolates was 80% and 100% of the biofilm formation in 46.87 and 93.75 µg/ml respectively. The methanolic and aqueous leaves extracts from Camellia sinensis are a successful method for producing CAgNPs. The synthesized CAgNPs also have significant antibacterial activity against S. aureus, depending on the concentrations, and inhibit S. aureus biofilm formation.

Comparative analysis of chloroplast genome of Lonicera japonica cv. Damaohua.

Lonicera japonica is a well-known medicinal plant, and the Damaohua cultivar is one of the oldest known honeysuckle cultivars in China. The 155,151 bp chloroplast genome of this cultivar was obtained through Illumina sequencing. The genome includes a pair of inverted repeats (IRa and IRb; 23,789 bp each), a large single-copy region (88,924 bp), and a small single-copy (SSC) region (18,649 bp). In total, 127 unique genes were identified: 80 protein-coding, 39 tRNA, and 8 rRNA genes. Only ycf3 contained two introns. Eighty-nine large repetitive sequences and 54 simple sequence repeats were detected. Fifty potential RNA editing sites were predicted. Adaptive evolution analysis revealed that infA, matK, petB, petD, rbcL, rpl16, rpl2, rps3, ycf1, and ycf2 were positively selected, possibly reflecting the specific environmental adaptations of this cultivar. Sequence alignment and analysis revealed several candidate fragments for Lonicera species identification, such as the intergenic regions rpoB-petN, rbcL-accD, and psaA-ycf3. The IR region boundary and phylogenetic analysis revealed that the L. japonica cv. Damaohua chloroplast genome was most closely related to the L. japonica genome, but there were five distinct differences between the two. There are four sites with high variability between L. japonica and L. japonica cv. Damaohua with nucleotide variability (Pi) greater than 0.002, including rps2-rpoC2, atpB-rbcL, ycf1, and ycf1-trnN GUU. The differences between L. japonica and L. japonica cv. Damaohua were further confirmed by the single nucleotide polymorphism sites between these two species. Therefore, this study revealed that the chloroplast genome can serve as a universal super barcode for plant identification, which can identify differences and help distinguish Lonicera japonica from related species. An understanding of Lonicera japonica cv. Damaohua chloroplast genomics and a comparative analysis of Lonicera species will provide a scientific basis for breeding, species identification, systematic evolution analysis, and chloroplast genetic engineering research on medicinal honeysuckle plants.