Stable formulations for RNA and plasmid DNA are a matter of paramount significance in several fields, ranging from medicine to biotechnology. We have investigated the potential of 15 compounds derived from natural osmolytes to enhance the thermostability and protection of both RNA and plasmid DNA. Our findings demonstrated that several compounds exhibit remarkable effects, enhancing the long-term storage of plasmid DNA at room temperature and the resilience of RNA to high-temperature stress, surpassing the performance of commercial osmolytes. Importantly, we found that one of the compounds enhanced the detection efficacy of a cost-effective RT-PCR test for COVID-19 that we had previously developed. This work offers new possibilities for expanding the capabilities of molecular diagnostic assays and nucleic acid storage methods.
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http://dx.doi.org/10.1002/biot.202400608 | DOI Listing |
Hlife
January 2025
Tumor Virus RNA Biology Section, HIV Dynamics and Replication Program, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Maryland, USA.
Plasmid DNA transfection is one of the fundamental tools of biomedical research. Here, we found that plasmid DNA transfection mediated by liposomes activates multiple innate immune responses in several widely used cell lines. Their activations were visible by detection of stress granules (SG) and cGAS-DNA condensates (cGC) in the transfected cells in a plasmid DNA dose-dependent manner.
View Article and Find Full Text PDFInt J Mol Sci
February 2025
School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
Due to the barriers imposed by the restriction-modification (RM) system, Nisin-producing industrial strains of often encounter low transformation efficiency, which seriously hinders the widespread application of genetic engineering in non-model . Herein, we present a novel pre-modification strategy (PMS) coupled with optimized plasmid delivery systems designed to systematically evade RM barriers and substantially improve Nisin biosynthesis in . Through the use of engineered strains with methylation profiles specifically optimized for C20, we have effectively evaded RM barriers, thereby facilitating the efficient introduction of large Nisin biosynthetic gene clusters into .
View Article and Find Full Text PDFInt J Mol Sci
February 2025
Sino-French Institute of Nuclear Engineering and Technology, Sun Yat-sen University, Zhuhai 519082, China.
Studying different types of DNA damage induced by ultra-high-dose-rate (UHDR) irradiation is essential for understanding the mechanism underlying the FLASH effect. pBR322 plasmid DNA was irradiated using an electron FLASH beam. The content of each subtype of plasmid DNA was measured via gel electrophoresis, and the extent of DNA double-strand breaks (DSBs) and single-strand breaks (SSBs) under UHDR and conventional-dose-rate irradiation (CONV) was quantitatively compared.
View Article and Find Full Text PDFAnimals (Basel)
March 2025
Department of Companion Animals Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10230, Thailand.
A conventional PCR (cPCR) remains an effective molecular technique for the diagnosis of canine monocytic ehrlichiosis. However, agarose gel electrophoresis requires additional time after thermal cycling. In the present study, we developed a PCR-based lateral flow biosensor (PCR-LFB) to detect ().
View Article and Find Full Text PDFAnimals (Basel)
March 2025
Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400715, China.
Deciphering the gastrointestinal microbial response to oral SS DNA vaccines with different doses is helpful for identifying the mechanism for effective utilization of the vaccine for improving animal production. Here, we conduct a comparative study with different doses of vaccine (control: empty plasmid; low dose: 1 × 10 CFU vaccine; high dose: 1 × 10 CFU vaccine) using goat as a case to investigate the potential of somatostatin vaccination from the entire gastrointestinal microbiota perspective. Our results show that body weight gain and slaughter rate are greater in the L_SS group than in the C_SS group.
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