Transfection Protocol
Transfection Protocol - Related Videos
08:08
Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation
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Cited by 11 •
2011
We present our optimized high-throughput nucleofection protocol as an efficient way of transfecting primary human monocyte-derived dendritic cells with either plasmid DNA or siRNA without causing cell maturation. We further provide evidence for successful siRNA silencing of targeted gene RIG-I at both the mRNA and protein levels.
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...
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...
04:21
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.
11:24
Transfecting Human Neural Stem Cells with the Amaxa Nucleofector
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Cited by 10 •
2007
Introducing a gene of interest into a cell is a powerful method for elucidating its function in vivo. This protocol describes an efficient method of transfecting a culture of human neural stem/precursor cells (hNSPCs) using the Nucleofector electroporation apparatus made by Amaxa.
FAQs
These delivery approaches use different ways to help nucleic acids cross the cell membrane. Chemical reagents and lipid-based carriers associate with the genetic material and support cellular entry, whereas electroporation uses an applied electrical treatment to facilitate passage across the membrane. The appropriate choice depends on the cell type, the nucleic acid being delivered, and the balance between delivery efficiency and cellular toxicity.
Experimental design must account for whether the introduced material needs to reach the cytoplasm or the nucleus. This destination influences which delivery approach and cell conditions are appropriate. Because cell type affects how material enters and is released within cells, researchers optimize the protocol around the desired biological outcome, such as changing gene expression or producing a protein.
Cell density, reagent concentration, exposure time, and recovery conditions are central variables. Increasing delivery conditions without considering cell health can raise toxicity, while insufficient exposure or unsuitable reagent levels may reduce the amount of genetic material entering cells. Systematically adjusting these factors helps researchers identify conditions that provide effective delivery while preserving viable cells for later analysis.
The nucleic acid selected should match the intended biological result. DNA can support gene expression or recombinant protein production, while RNA and small interfering RNA can be used to alter gene expression in different experimental designs. This choice connects the delivery step to the study objective, whether researchers are examining gene function, modifying expression, or modeling a disease-related process.
A practical optimization plan varies cell density, reagent concentration, exposure time, and recovery conditions while monitoring both delivery efficiency and cellular toxicity. Researchers can compare how these variables affect the desired outcome, then select conditions that maintain an appropriate balance. This controlled adjustment is important because a protocol that works for one cell type or design may not perform similarly in another.
Transfection supports several experimental goals, including gene function studies, recombinant protein production, genome editing, and disease modeling. It allows researchers to connect introduced nucleic acids with measurable changes in gene expression or protein output. The method is therefore useful both for investigating how genes operate and for creating cellular systems that represent particular biological or disease-related conditions.