Cell Transfection
Cell Transfection - Related Videos
10:24
Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells
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Cited by 26 •
2011
Despite recent advancements in genetic modification, transfection of human embryonic stem cells (HESCs) remains a capricious process. To our knowledge, systematic and efficient methods to transfect human induced pluripotent stem cells (iPSCs) have not been reported. Here, we describe robust protocols to efficiently transfect and nucleofect human iPSCs.
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...
08:46
Single Cell Transfection in Chick Embryos
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Cited by 3 •
2010
Using fine tip micropipettes we inject plasmid DNA into subdomains of chicken somites or neural tubes. The concentration of the plasmid is adjusted to generate single transfected cells. We then allow the cells to develop into clonal populations.
12:58
Transfecting RAW264.7 Cells with a Luciferase Reporter Gene
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Cited by 13 •
2015
Transfection into the macrophage cell line, RAW264.7, is difficult due to the cell’s natural response against foreign materials. We described here a gentle yet robust procedure for transfecting luciferase reporter genes into RAW264.7 cells.
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.
FAQs
Lipid-based carriers form complexes with nucleic acids and help them cross the cell membrane. This approach can deliver DNA, mRNA, or small interfering RNA into living cells, allowing researchers to alter gene expression or produce specific proteins. The selected nucleic acid and carrier therefore influence whether the experiment focuses on gene-function analysis, protein production, or RNA interference.
Electroporation applies brief electrical pulses that create temporary pores in the cell membrane. These openings provide a route for nucleic acids to enter living cells without relying on lipid-based complexes. Because the pores are temporary, the electrical treatment is used as a distinct delivery mechanism for experiments requiring introduced DNA, mRNA, or small interfering RNA.
Transient transfection supports short-term analysis of cellular responses after nucleic acid delivery, whereas stable transfection is intended to maintain introduced genetic material across cell divisions. This distinction determines the experimental time frame: transient approaches suit immediate gene-expression or protein-production studies, while stable approaches support analyses that require persistence through repeated cell division.
The nucleic acid cargo determines the type of biological change being investigated. DNA can be introduced to support gene-expression studies or production of a specific protein, mRNA can be used when protein production is the focus, and small interfering RNA can alter gene expression through RNA interference. Matching the cargo to the question keeps the experiment biologically focused.
A typical experiment selects a nucleic acid cargo, chooses either lipid-based delivery or electroporation, introduces the material into living cells, and then examines the resulting response. The readout may involve altered gene expression, production of a specific protein, or another application such as a reporter assay. The workflow therefore connects delivery method with the intended biological outcome.
Reporter assays are useful when researchers need to examine the consequences of introduced genetic material through a defined experimental readout. Cell Transfection enables DNA, mRNA, or small interfering RNA to be delivered before the assay, allowing investigators to study changes associated with gene expression or regulation. This makes transfection a practical component of controlled gene-function experiments.
For RNA interference, transfection delivers small interfering RNA into living cells to alter gene expression and examine the resulting biological effects. It can also serve as part of genome-editing workflows by introducing the nucleic acids required for that experimental approach. In both cases, transfection provides the delivery step that connects selected genetic material with a targeted biology question.