Reverse Transfection
Reverse Transfection - Related Videos
08:04
Neuronal Transfection Methods
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2023
Transfection - the process of transferring genetic material into cells - is a powerful tool for the rapid and efficient manipulation of gene expression in cells. Because this method can be used to silence the expression of specific proteins or to drive the expression of foreign or modified proteins, transfection is an extremely useful tool in the study of the cellular and molecular processes that govern neuron function. However, mature neurons have a number of properties that make them...
04:34
siRNA Reverse Transfection-Based Gene Silencing In Vitro: A Procedure to Deliver Small Interfering RNA in Cultured Cells to Inactivate Target Gene Expression
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2025
In this video, we describe a procedure to perform reverse transfection of siRNA into adipocytes to modulate the expression of a target protein.
07:28
An Introduction to Transfection
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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...
04:21
Protein Transfection of Mouse Lung
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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.
08:21
Time-lapse Imaging of Mitosis After siRNA Transfection
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Cited by 8 •
2010
Here we describe a basic protocol to image and quantify the mitotic timing of live mammalian tissue culture cells after siRNA transfection.
FAQs
The cargo directs the type of cellular response measured after delivery. DNA can support studies of gene function and protein production, messenger RNA can be used to regulate protein production, and small interfering RNA can produce gene silencing. Selecting among these cargos therefore connects the experimental design to expression, suppression, or pathway-related questions.
Lipid or polymer carriers are combined with nucleic acids to form transfection complexes before cells are introduced. These complexes provide the material that cells subsequently take up, commonly through endocytosis, a process in which cells internalize external material. Carrier-based complex formation is therefore central to moving the nucleic acid into cultured cells for downstream analysis.
Endocytosis represents a common uptake route for the complexes by cultured cells. After uptake, the delivered nucleic acid can produce the intended downstream effect, such as gene expression or silencing, depending on the cargo. This link between internalization and cellular response helps explain why the experiment is evaluated through changes in protein production, gene function, or pathway behavior.
A basic workflow begins by combining DNA, messenger RNA, or small interfering RNA with a lipid or polymer carrier in the culture vessel. Cells are then seeded onto the prepared complexes and maintained as a cultured-cell experiment. Subsequent measurements focus on the resulting gene expression, protein production, silencing, or other biological response relevant to the study.
This format is especially useful when many conditions must be processed in parallel. Preparing complexes in the culture vessel before cell seeding reduces handling steps and supports parallel processing, making the approach suitable for high-throughput screening. Such experiments can compare multiple nucleic acid cargos or gene-related conditions while monitoring cellular responses across cultured-cell samples.
In biology, the method can support gene-function studies, regulation of protein production, and modeling of cellular pathways. It can also be used to evaluate candidate therapeutics in cultured cells. The measured outcome depends on the selected nucleic acid, allowing experiments to examine either increased gene-related activity or targeted silencing within a defined cellular system.