Ribonucleoprotein Electroporation
Ribonucleoprotein Electroporation - Related Videos
08:31
CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
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Cited by 5 •
2019
Presented here is a protocol for efficient CRISPR/Cas9 ribonucleoprotein-mediated gene editing in mammalian cells using tube electroporation.
09:45
Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins and mRNA into Freshly Isolated Primary Mouse Hepatocytes
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Cited by 3 •
2022
This protocol describes techniques for isolating primary mouse hepatocytes from the liver and electroporating CRISPR-Cas9 as ribonucleoproteins and mRNA to disrupt a therapeutic target gene associated with an inherited metabolic disease of the liver. The methods described result in high viability and high levels of gene modification after electroporation.
11:57
Electroporation of Mycobacteria
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Cited by 55 •
2008
Mycobacterial pathogenic strategies remain poorly understood. The slow growth rate of most species, the impenetrable nature of the cell-wall, and the hazards of working with pathogens make mycobacteria difficult to study and are largely responsible for our poor understanding of these organisms. In this video we will demonstrate the technique of electroporation, which involves subjecting cells to a brief high electrical impulse to allow the entry of DNA. It is the most widely used method for...
08:05
Murine In Utero Electroporation
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2023
In utero electroporation is an important technique for studying the molecular mechanisms that guide the proliferation, differentiation, migration, and maturation of cells during neural development. Electroporation enables the rapid and targeted delivery of material into cells by utilizing electrical pulses to create transient pores in cell membranes. Although electroporation has traditionally been used in in vitro studies, scientific advancements have now broadened its utilization to intact...
12:19
Bacterial Transformation: Electroporation
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2023
The term “transformation” refers cellular ingestion of foreign DNA. In nature, transformation can occur in certain types of bacteria. In molecular biology, however, transformation is artificially induced through the creation of pores in the bacterial cell walls. Bacterial cells that are able to take up DNA from the environment are called competent cells. Electrocompetent cells can be produced in the laboratory and transformation of these cells can be achieve via the application of an...
FAQs
Brief electrical pulses temporarily permeabilize the cell membrane, creating an opportunity for preassembled Cas9–guide RNA complexes to cross into the cell. Once the pulse has passed, the membrane can recover while the complexes continue toward the cytoplasm and nucleus. This temporary permeability is central to delivering editing reagents without introducing DNA into the cell.
The guide RNA directs Cas9 to a specific DNA sequence, where the nuclease creates a targeted break. Cellular repair of that break can produce insertions or deletions, supporting gene knockout or functional disruption. If a donor template is supplied, repair can instead support precise sequence replacement, changing the type of genetic outcome produced.
The editing components enter cells as preassembled protein–RNA complexes rather than as persistent DNA-encoding material. Their transient exposure supports rapid editing while reducing the risk of vector integration. This distinction is useful when researchers want targeted genetic modification without maintaining a delivered DNA construct as part of the experimental system.
A typical workflow begins by assembling Cas9 with a guide RNA and preparing the target cells for electrical delivery. The cell suspension then receives brief electrical pulses that permit complex entry. Afterward, cellular repair processes the Cas9-induced DNA break. Researchers may include a donor template when the experiment requires precise sequence replacement rather than insertion or deletion outcomes.
This approach is suited to rapid gene knockout experiments, functional screening, and production of engineered cells. Because the editing reagent is delivered as a protein–RNA complex, researchers can directly test targeted genetic changes without relying on DNA delivery. The method therefore supports experiments that prioritize targeted modification, transient exposure, and reduced vector-integration risk.
The outcome depends on how cells repair the targeted DNA break. Insertions or deletions indicate repair-associated sequence disruption and can support knockout studies, whereas incorporation of a supplied donor template indicates a route toward precise replacement. Comparing these outcome types helps researchers distinguish functional gene disruption from intentional sequence modification in engineered cells.