EGFP (enhanced green fluorescent protein) is one of the most common tools used in life sciences, including research focusing on proteostasis. Here we report that ERN1 (endoplasmic reticulum to nucleus signaling 1), which is upregulated by UPR (unfolded protein response), targets an RNA hairpin loop motif in EGFP mRNA. A silent mutation introduced into EGFP mRNA abolished the ERN1-dependent mRNA decay. Therefore, experiments that employ EGFP as a reporter gene in studies that involve upregulation of the UPR pathway should be interpreted carefully, and a mutant devoid of the ERN1 target motif may be more suitable for such studies.
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http://dx.doi.org/10.1002/jcb.30314 | DOI Listing |
Adv Sci (Weinh)
January 2025
Department of Internal Medicine III, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074, Aachen, Germany.
Most gene therapies exert their actions via manipulation of hepatocytes (parenchymal cells) and the reasons behind the suboptimal performance of synthetic mRNA in non-parenchymal cells (NPC) such as Kupffer cells (KC), and liver macrophages, remain unclear. Here, the spatio-temporal distribution of mRNA encoding enhanced green fluorescent protein (Egfp), siRNA, or both co-encapsulated into lipid nanoparticles (LNP) in the liver in vivo using real-time intravital imaging is investigated. Although both KC and hepatocytes demonstrate comparable high and rapid uptake of mRNA-LNP and siRNA-LNP in vivo, the translation of Egfp mRNA occurs exclusively in hepatocytes during intravital imaging.
View Article and Find Full Text PDFEnviron Res
January 2025
Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China. Electronic address:
Hygienic insecticides are applied directly to the living environment and are closely related to human life. Dimefluthrin (DIM) is one of the most widely used hygienic insecticides globally. However, with increasing mosquito resistance, both the concentration and duration of DIM usage have risen, prompting public concerns regarding its neurotoxic risks, especially for immunocompromised children.
View Article and Find Full Text PDFAs an advanced nucleic acid therapeutical modality, mRNA can express any type of protein in principle and thus holds great potential to prevent and treat various diseases. Despite the success in COVID-19 mRNA vaccines, direct local delivery of mRNA into the lung by inhalation would greatly reinforce the treatment of pulmonary pathogens and diseases. Herein, we developed lipid nanoparticles (LNPs) from degradable ionizable glycerolipids for potent pulmonary mRNA delivery via nebulization.
View Article and Find Full Text PDFFront Behav Neurosci
December 2024
Department of Psychology, University of California, Davis, Davis, CA, United States.
Molecular and genetic techniques now allow selective tagging and manipulation of the population of neurons, often referred to as "engram cells," that were active during a specific experience. One common approach to labeling these cells is to use the transgenic mouse (TetTag). In addition to tagging cells active during learning, it is common to examine the reactivation of these cells using immediate early gene (IEG) expression as an index of neural activity.
View Article and Find Full Text PDFBiomed Rep
February 2025
Department of Molecular Pharmaceutics, Hoshi University, Shinagawa, Tokyo 142-8501, Japan.
Previously, it was reported that mRNA/cationic liposome complexes (mRNA lipoplexes) composed of the cationic triacyl lipid, 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl)propan-2-yl)amino)-,,- trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12), with 1,2-dioleoyl-glycero-3-phosphoethanolamine and poly(ethylene glycol) cholesteryl ether, induce high protein expression in human cervical carcinoma HeLa cells. In the present study, the authors aimed to optimize mRNA transfection using TC-1-12-based mRNA lipoplexes. mRNA lipoplexes were prepared at various charge ratios (+:-) using modified ethanol injection (MEI) and thin-film hydration (TFH) methods and compared the protein expression efficiency after transfection of HeLa cells with the developed mRNA lipoplexes.
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