Corrigendum to “Missing pieces in understanding the intracellular trafficking of polycation/DNA complexes” [J. Control. Release 139 (2) (2009) 88–93]

2011 ◽  
Vol 152 (2) ◽  
pp. 326 ◽  
Author(s):  
You-Yeon Won ◽  
Rahul Sharma ◽  
Stephen F. Konieczny
Keyword(s):  
Intracellular Trafficking ◽  
Control Release ◽  
Dna Complexes

scholarly journals SNAPSwitch: A Molecular Sensor to Quantify the Localization of Proteins, DNA and Nanoparticles in Cells

2019 ◽  
Author(s):  
Laura I FitzGerald ◽  
Luigi Aurelio ◽  
Moore Chen ◽  
Daniel Yuen ◽  
Bim Graham ◽  
...  
Keyword(s):  
Immune Responses ◽  
Intracellular Trafficking ◽  
Live Cells ◽  
Quantitative Detection ◽  
Dna Complexes ◽  
Receptor Signalling ◽  
Fluorescent Markers ◽  
Molecular Sensor ◽  
Protein Nucleic Acid ◽  
In Cells

Intracellular trafficking governs receptor signalling, pathogenesis, immune responses and the cellular fate of nanomedicines. These processes are typically tracked by confocal microscopy, where colocalization of fluorescent markers implies an interaction or co-compartmentalization. However, this type of analysis is inherently low-throughput, is limited by the resolution of microscopy, and can miss fleeting interactions. To address this, we have developed a localization sensor composed of a quenched and attachable SNAP-tag substrate (SNAPSwitch). SNAPSwitch enables quantitative detection of protein, nucleic acid and nanoparticle trafficking to locations of interest within live cells using flow cytometry. Using this approach, we followed the trafficking of DNA complexes travelling from endosomes into the cytosol and to the nucleus. We also show that antibody targeted to the transferrin (CD71) or hyaluronan (CD44) receptor is initially sorted into different compartments following endocytosis. These results demonstrate SNAPSwitch is a high-throughput and broadly applicable tool to quantitatively track the localization of materials in cells.

scholarly journals Binding Affinity, Cellular Uptake, and Subsequent Intracellular Trafficking of the Nano-Gene Vector P123-PEI-R13

2016 ◽  
Vol 2016 ◽  
pp. 1-8
Author(s):  
Yaguang Zhang ◽  
Hongmei Shu ◽  
Jing Hu ◽  
Min Zhang ◽  
Junweng Wu ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Cellular Uptake ◽  
Intracellular Trafficking ◽  
Transfection Efficiency ◽  
Cross Linking ◽  
Low Molecular Weight ◽  
Gene Vector ◽  
Dna Complexes ◽  
Dynamic Source

A nano-gene vector PEI-P123-R13 was synthesized by cross-linking low molecular weight PEI with P123 and further coupling bifunctional peptide R13 to the polymer for targeting tumor and increasing cellular uptake. The binding assessment of R13 toαvβ3 positive cells was performed by HRP labeling. The internalization pathways of P123-PEI-R13/DNA complexes were investigated based on the effect of specific endocytic inhibitors on transfection efficiency. The mechanism of intracellular trafficking was investigated based on the effect of endosome-lysosome acidification inhibitors, cytoskeleton, and dynein inhibitors on transfection efficiency. The results indicated that the bifunctional peptide R13 had the ability of binding toαvβ3 positive cellsin vitro. The modification of P123-PEI-R13 with R13 made it display new property of internalization. P123-PEI-R13/DNA complexes were conducted simultaneously via clathrin-mediated endocytosis, caveolin-mediated endocytosis, macropinocytosis, and possible energy-independent route. After internalization, P123-PEI-R13/DNA complexes could escape from the endosome-lysosome system because of its acidification and further took microtubule as the track and dynein as the dynamic source to be transported toward the microtubule (+) end, to wit nucleus, under the action of microfilament, and with the aid of intermediate filament.

scholarly journals Intracellular trafficking of cationic liposome–DNA complexes in living cells

Soft Matter ◽  
2012 ◽  
Vol 8 (30) ◽  
pp. 7919 ◽  
Author(s):  
Stefano Coppola ◽  
Laura C. Estrada ◽  
Michelle A. Digman ◽  
Daniela Pozzi ◽  
Francesco Cardarelli ◽  
...  
Keyword(s):  
Intracellular Trafficking ◽  
Cationic Liposome ◽  
Living Cells ◽  
Dna Complexes

scholarly journals Missing pieces in understanding the intracellular trafficking of polycation/DNA complexes

2009 ◽  
Vol 139 (2) ◽  
pp. 88-93 ◽  
Author(s):  
You-Yeon Won ◽  
Rahul Sharma ◽  
Stephen F. Konieczny
Keyword(s):  
Intracellular Trafficking ◽  
Dna Complexes

scholarly journals A molecular sensor to quantify the localization of proteins, DNA and nanoparticles in cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Laura I. FitzGerald ◽  
Luigi Aurelio ◽  
Moore Chen ◽  
Daniel Yuen ◽  
Joshua J. Rennick ◽  
...  
Keyword(s):  
Immune Responses ◽  
Intracellular Trafficking ◽  
Cellular Membrane ◽  
Live Cells ◽  
Quantitative Detection ◽  
Membrane Material ◽  
Dna Complexes ◽  
Fluorescent Markers ◽  
Molecular Sensor ◽  
In Cells

Abstract Intracellular trafficking governs receptor signaling, pathogenesis, immune responses and fate of nanomedicines. These processes are typically tracked by observing colocalization of fluorescent markers using confocal microscopy. However, this method is low throughput, limited by the resolution of microscopy, and can miss fleeting interactions. To address this, we developed a localization sensor composed of a quenched SNAP-tag substrate (SNAPSwitch) that can be conjugated to biomolecules using click chemistry. SNAPSwitch enables quantitative detection of trafficking to locations of interest within live cells using flow cytometry. Using SNAPSwitch, we followed the trafficking of DNA complexes from endosomes into the cytosol and nucleus. We show that antibodies against the transferrin or hyaluronan receptor are initially sorted into different compartments following endocytosis. In addition, we can resolve which side of the cellular membrane material was located. These results demonstrate SNAPSwitch is a high-throughput and broadly applicable tool to quantitatively track localization of materials in cells.

Torus Circumference and Thickness Parameters From a Linear DNA Size Series Suggest How Toruses Self-Assemble

1985 ◽  
Vol 43 ◽  
pp. 522-523
Author(s):  
George C. Ruben ◽  
Kenneth A. Marx
Keyword(s):  
Tertiary Structure ◽  
Dna Complexes ◽  
Tertiary Structures ◽  
Self Assembling ◽  
Double Stranded Dna ◽  
Calf Thymus ◽  
Dna Organization ◽  
Condensed Dna ◽  
Dna Size

Certain double stranded DNA bacteriophage and viruses are thought to have their DNA organized into large torus shaped structures. Morphologically, these poorly understood biological DNA tertiary structures resemble spermidine-condensed DNA complexes formed in vitro in the total absence of other macromolecules normally synthesized by the pathogens for the purpose of their own DNA packaging. Therefore, we have studied the tertiary structure of these self-assembling torus shaped spermidine- DNA complexes in a series of reports. Using freeze-etch, low Pt-C metal (10-15Å) replicas, we have visualized the microscopic DNA organization of both calf Thymus( CT) and linear 0X-174 RFII DNA toruses. In these structures DNA is circumferentially wound, continuously, around the torus into a semi-crystalline, hexagonal packed array of parallel DNA helix sections.

Small GTP-binding Protein RalA Associates with Weibel-Palade Bodies in Endothelial Cells

1999 ◽  
Vol 82 (09) ◽  
pp. 1177-1181 ◽  
Author(s):  
Hubert de Leeuw ◽  
Pauline Wijers-Koster ◽  
Jan van Mourik ◽  
Jan Voorberg
Keyword(s):  
Endothelial Cells ◽  
Binding Proteins ◽  
Binding Protein ◽  
Control Release ◽  
Small Gtpase ◽  
Gtp Binding Protein ◽  
Two Dimensional Gel Electrophoresis ◽  
Gtp Binding Proteins ◽  
Dense Granules ◽  
Gtp Binding

SummaryIn endothelial cells von Willebrand factor (vWF) and P-selectin are stored in dense granules, so-called Weibel-Palade bodies. Upon stimulation of endothelial cells with a variety of agents including thrombin, these organelles fuse with the plasma membrane and release their content. Small GTP-binding proteins have been shown to control release from intracellular storage pools in a number of cells. In this study we have investigated whether small GTP-binding proteins are associated with Weibel-Palade bodies. We isolated Weibel-Palade bodies by centrifugation on two consecutive density gradients of Percoll. The dense fraction in which these subcellular organelles were highly enriched, was analysed by SDS-PAGE followed by GTP overlay. A distinct band with an apparent molecular weight of 28,000 was observed. Two-dimensional gel electrophoresis followed by GTP overlay revealed the presence of a single small GTP-binding protein with an isoelectric point of 7.1. A monoclonal antibody directed against RalA showed reactivity with the small GTP-binding protein present in subcellular fractions that contain Weibel-Palade bodies. The small GTPase RalA was previously identified on dense granules of platelets and on synaptic vesicles in nerve terminals. Our observations suggest that RalA serves a role in regulated exocytosis of Weibel-Palade bodies in endothelial cells.

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