چکیده مقاله
This comprehensive study elucidates the de novo synthesis and physicochemical fine tuning of hierarchically engineered multilaminar nanocarrier assemblies encapsulating CRISPR/Cas9 ribonucleoprotein RNP complexes, devised for ultra precise, locus specific genomic perturbation of essential biofilm regulatory operons within recalcitrant bacterial biofilm consortia prevalent in agro industrial food processing ecosystems Fabrication employed iterative layer by layer LbL polyelectrolyte self assembly integrated with surface stealthing via polyethylene glycol PEG conjugation and targeting ligand functionalization to optimize endocytic uptake kinetics, circumvent intracellular endonuclease mediated degradation, and mediate temporospatial endosomal escape culminating in efficacious cytoplasmic RNP bioavailability Highly specific, computationally optimized single guide RNAs sgRNAs were strategically designed to target conserved genetic loci responsible for exopolysaccharide EPS biosynthesis pathways, quorum sensing hierarchical networks, c di GMP signaling cascades, and multidrug resistance MDR efflux systems, all critically orchestrating biofilm maturation, extracellular polymeric substance EPS matrix robustness, and phenotypic antibiotic tolerance Post delivery, extensive transcriptomic rewiring was quantitatively profiled via ultra deep, strand specific RNA sequencing RNA Seq , employing stringent normalization and differential expression models complemented by integrative systems biology platforms including gene set enrichment analysis GSEA , weighted gene co expression network analysis WGCNA , and regulatory motif discovery to elucidate disruption of transcriptional modules underpinning biofilm resilience and antimicrobial defense Functional validation encompassing confocal laser scanning microscopy CLSM and high throughput crystal violet biofilm biomass quantification assays confirmed significant attenuation of biofilm architectural integrity, EPS matrix deposition, and MDR phenotype suppression, thereby substantiating the genomic editing efficacy The integrative theranostic nano genomic platform described herein exemplifies a revolutionary modality for precision microbial biofilm modulation, enabling programmable, scalable, and biosafe biocontrol interventions in industrial food production lines, advancing microbial ecology engineering frontiers, enhancing product sterility, and mitigating entrenched biofilm driven contamination risks with far reaching implications for next generation antimicrobial therapeutics and food safety technologies
کلیدواژهها
نویسندگان
Behzad beizaei
Master's student in Food Science and Engineering, Islamic Azad University, Tehran Science and Research Branch
Morteza Jamshid Eini
PhD in Food Technology, Islamic Azad University, North Tehran Branch
شیوه ارجاع
beizaei, Behzad and jamshid eini, Morteza,1404,CRISPR/Cas9-Loaded Nanocarriers for Targeted Editing of Biofilm-Regulatory Genes in Resistant Food Industry Strains with Advanced Quantitative RNA-Seq Transcriptomic Analysis,Ninth International Conference on Interdisciplinary Studies in Food industry and Nutrition Science of Iran,Tehran
ارائهشده در
مجموعه مقالات نهمین همایش بین المللی مطالعات میان رشته ای در صنایع غذایی و علوم تغذیه ایران29 آبان 1404 · تهران