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Long-term blood glucose control via glucose-activated transcriptional regulation of insulin analogue in type 1 diabetes mice.
Diabetes Obes Metab
January 2025
National Engineering Research Center for Biomaterials, College of Biomedical Engineering, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Aim: To achieve glucose-activated transcriptional regulation of insulin analogue in skeletal muscle of T1D mice, thereby controlling blood glucose levels and preventing or mitigating diabetes-related complications.
Materials And Methods: We developed the GANIT (Glucose-Activated NFAT-regulated INSA-F Transcription) system, an innovative platform building upon the previously established intramuscular plasmid DNA (pDNA) delivery and expression system. In the GANIT system, skeletal muscle cells are genetically engineered to endogenously produce the insulin analogue INSA-F (Insulin Aspart with Furin cleavage sites).
Metal-Coordinated Histidine-Functionalized Redox-Responsive Polyethyleneimine as a Smart Gene Delivery Vector.
Mol Biotechnol
January 2025
Noncommunicable Disease Research Center, Jahrom University of Medical Sciences, Jahrom, Iran.
Despite significant advancements in gene delivery and CRISPR technology, several challenges remain. Chief among these are overcoming serum inhibition and achieving high transfection efficiency with minimal cytotoxicity. To address these issues, there is a need for novel vectors that exhibit lower toxicity, maintain stability in serum-rich environments, and effectively deliver plasmids of various sizes across diverse cell types.
View Article and Find Full Text PDFPlasmid Stability Analysis with Open-Source Droplet Microfluidics.
J Vis Exp
December 2024
Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile;
Plasmids play a vital role in synthetic biology by enabling the introduction and expression of foreign genes in various organisms, thereby facilitating the construction of biological circuits and pathways within and between cell populations. For many applications, maintaining functional plasmids without antibiotic selection is critical. This study introduces an open-hardware-based microfluidic workflow for analyzing plasmid retention by culturing single cells in gel microdroplets and quantifying microcolonies using fluorescence microscopy.
View Article and Find Full Text PDFCryo-EM structure of the conjugation H-pilus reveals the cyclic nature of the TrhA pilin.
bioRxiv
December 2024
Rutherford Appleton Laboratory, Research Complex at Harwell, Didcot, Oxfordshire, UK.
Conjugation, the major driver of the spread of antimicrobial resistance genes, relies on a conjugation pilus for DNA transfer. Conjugative pili, such as the F-pilus, are dynamic tubular structures, composed of a polymerized pilin, that mediate the initial donor-recipient interactions, a process known as mating pair formation (MPF). IncH are low-copy-number plasmids, traditionally considered broad host range, which are found in bacteria infecting both humans and animals.
View Article and Find Full Text PDFSimple and Fail-safe Method to Transform Miniprep Strain K12 Plasmid DNA Into Viable EHA105 Cells for Plant Genetic Transformation.
Bio Protoc
January 2025
Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Nairobi, Kenya.
Agrobacterium-mediated gene transformation method is a vital molecular biology technique employed to develop transgenic plants. Plants are genetically engineered to develop disease-free varieties, knock out unsettling traits for crop improvement, or incorporate an antigenic protein to make the plant a green factory for edible vaccines. The method's robustness was validated through successful transformations, demonstrating its effectiveness as a standard approach for researchers working in plant biotechnology.
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