Organoid Transfection
Organoid Transfection - Related Videos
05:11
Organoid Electroporation: A Method to Transfect Plasmid DNA into Gastrointestinal Organoids
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2023
This video describes the electroporation protocol for transfecting large plasmids into three-dimensional gastrointestinal organoids to prove its universal functionality in different entities, namely pancreatic ductal adenocarcinoma (PDAC), colorectal cancer (CRC), cholangiocarcinoma (CCC), and gastric cancer (GC) organoids.
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...
10:58
A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids
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Cited by 81 •
2015
In this video protocol we give a step by step explanation of lentiviral transduction in organoids of primary intestinal epithelium and of processing and downstream analysis of these cultures by quantitative RT-PCR, RNA-microarray and immunohistochemistry.
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.
FAQs
Delivery depends on how nucleic acids cross the membranes of organoid cells. Lipid-based carriers and electroporation provide alternative ways to facilitate entry, allowing researchers to introduce DNA, RNA, or genome-editing reagents. This choice is important because the delivered material determines whether the experiment is designed for temporary genetic activity or for targeted modification of cellular genomes.
The introduced cargo largely determines the outcome. DNA or RNA can support transient expression, meaning genetic activity occurs without necessarily producing a permanent genome change. Genome-editing reagents, in contrast, can be used for targeted genetic modification. Distinguishing these outcomes helps researchers interpret whether an observed phenotype reflects temporary molecular activity or an altered genetic state.
Maintaining cellular organization allows genetic changes to be examined in a tissue-like setting rather than only as isolated molecular events. Researchers can therefore connect altered gene activity or genome sequence with effects on tissue-level phenotypes. This relationship is especially valuable when cellular interactions and organization contribute to how development, disease mechanisms, or treatment responses appear.
DNA, RNA, and genome-editing reagents serve different experimental purposes. DNA or RNA can produce temporary expression that helps test the effect of a gene or regulatory activity, while editing reagents support targeted genetic modification. Selecting among these cargos allows investigators to separate short-term functional tests from experiments intended to examine consequences of changed genetic information.
A study begins by selecting the genetic material and a delivery approach, such as a lipid-based carrier or electroporation. The material is introduced into organoid cells, followed by examination of the resulting expression, genetic modification, or tissue-level phenotype. This workflow connects a controlled molecular perturbation with outcomes relevant to gene function, development, disease, or treatment response.
The method supports gene-function studies, disease modeling, lineage tracing, and evaluation of regulatory elements. It can also be used to investigate therapeutic responses in a tissue-like context. These applications make organoids useful when researchers need to examine how a defined genetic manipulation influences organized cellular behavior rather than measuring molecular activity alone.
In genetics, the approach provides a way to manipulate organoid genomes while retaining aspects of tissue organization. Researchers can use that control to relate genes or regulatory elements to developmental patterns, disease-associated phenotypes, lineage behavior, and responses to therapies. The resulting connection between genetic change and tissue-level outcome strengthens interpretation of gene function in a human-tissue model.