Cargo Protein Transfection
Cargo Protein Transfection - Related Videos
08:06
Identification of Kinesin-1 Cargos Using Fluorescence Microscopy
0 Views •
Cited by 1 •
2016
Here, a protocol is presented to identify Kinesin-1 cargos. A motorless mutant of the Kinesin-1 heavy chain (KIF5B) aggregates in the cytoplasm and induces aggregation of its cargos. Both aggregates are detected under fluorescence microscopy. A similar strategy can be employed to identify cargos of other motor proteins.
04:21
Protein Transfection of Mouse Lung
0 Views •
Cited by 1 •
2013
Transgenic mice or viral vectors have been used to increase protein expression within the lung. However, these techniques are time-consuming, technically challenging and have off-target effects that can confound results. Our protein transfection protocol uses a lipid based transfection reagent and an ultrafine microsprayer to uniformly deliver active protein to lung cells.
02:35
Visualizing Intracellular Transport of Cargo in Cultured Astrocytes
0 Views •
2025
Source: Creighton, B. A., et al. Visualizing and Analyzing Intracellular Transport of Organelles and Other Cargos in Astrocytes. J. Vis. Exp. (2019)This video demonstrates the visualization of intracellular cargo transport in cultured astrocytes using a fluorescent acidotropic probe that selectively labels acidic endolysosomal vesicles. Time-lapse imaging and Z-stack acquisition with a confocal microscope allow tracking of fluorescently labeled cargo throughout the three-dimensional cell volume.
09:00
Cargo Loading onto Kinesin Powered Molecular Shuttles
0 Views •
Cited by 4 •
2010
Molecular shuttles consisting of functionalized microtubules gliding on surface-adhered kinesin motor proteins can serve as a nanoscale transport system. Here, the assembly of a typical shuttle system is described.
07:28
An Introduction to Transfection
0 Views •
2023
Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for transfected RNA is the cytoplasm, DNA must be transported to the nucleus for effective transfection. There, the...
FAQs
Carrier choice determines how a protein enters living cells. Cargo Protein Transfection may use cell-penetrating peptides, lipid-based reagents, nanoparticles, or physical delivery methods to cross the plasma membrane. After entry, the protein can act directly on intracellular targets instead of relying on gene expression, allowing experiments to examine cellular effects soon after delivery.
The delivered protein remains active only until the cell degrades it, creating a temporary period of manipulation. This differs from approaches that introduce DNA or depend on gene expression, and it avoids genomic integration. Researchers can therefore study rapid cellular responses while limiting the intervention to a defined, nonpermanent window.
Protein degradation limits how long the introduced activity persists inside a cell. Consequently, observed changes reflect a temporary intervention rather than a continuously maintained genetic program. This feature is important when interpreting cellular behavior, because measurements should be related to the period during which the cargo remains functional and able to act on intracellular targets.
A basic experiment combines a functional protein cargo with a delivery strategy capable of crossing the plasma membrane of living cells. Suitable options include cell-penetrating peptides, lipid-based reagents, nanoparticles, or physical delivery methods. After introduction, researchers assess the resulting cellular behavior while the protein remains active, before its eventual degradation limits the effect.
In neuroscience, this approach is useful when researchers need acute control over neuronal processes. Delivered proteins can be used to examine signaling, cytoskeletal dynamics, transcriptional regulation, or synaptic function without introducing DNA. The transient response makes the method suitable for studying rapid changes in neurons and other neural models.
Protein delivery can reveal how directly manipulated intracellular targets influence neuronal signaling, structural dynamics, transcriptional regulation, or synaptic function. Because the intervention does not rely on gene expression and remains temporary, researchers can connect these outcomes to an acute change in protein activity. The same strategy also supports testing protein-based interventions in neural models.