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Related Concept Videos

Transformation01:26

Transformation

Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...

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Electroporation of Functional Bacterial Effectors into Mammalian Cells
08:39

Electroporation of Functional Bacterial Effectors into Mammalian Cells

Published on: January 19, 2015

Transfection by electroporation.

Huntington Potter1, Richard Heller

  • 1Byrd Alzheimer's Institute, University of South Florida College of Medicine, Tampa, Florida, USA.

Current Protocols in Molecular Biology
|October 5, 2010
PubMed
Summary
This summary is machine-generated.

Electroporation uses electric shocks to deliver DNA into cells, offering an easier, efficient method for stable transformation and gene expression across various cell types. This technique is valuable for creating genetically modified mice, gene therapy, and DNA vaccination.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Cell Biology

Background:

  • Electroporation is a physical method for introducing molecules into cells.
  • It utilizes high-voltage electric pulses to permeabilize cell membranes.
  • This technique offers advantages over other transfection methods due to its efficiency and versatility.

Purpose of the Study:

  • To describe the application of electroporation for mammalian cell transfection.
  • To detail protocols for generating genetically modified mice (knock-out, knock-in, transgenic) using electroporation of ES cells.
  • To present in vivo electroporation methods for gene therapy and DNA vaccination, and plant protoplast transfection.

Main Methods:

  • High-voltage electric shocks applied to cells to induce DNA uptake.
  • Electroporation of mouse embryonic stem (ES) cells for genetic modification.
  • In vivo electroporation for therapeutic applications and vaccination.
  • Modification of protocols for plant protoplast preparation and transfection.

Main Results:

  • Electroporation is effective with most cell types.
  • Achieves high frequencies of stable transformation and transient gene expression.
  • Enables creation of knock-out, knock-in, and transgenic mice.
  • Demonstrates utility in in vivo gene therapy, cancer therapy, and DNA vaccination.
  • Adaptable for plant protoplast transfection.

Conclusions:

  • Electroporation is a versatile and efficient method for genetic material delivery into diverse cell types.
  • It is a key technique for generating genetically engineered animal models and advancing gene-based therapies.
  • The described protocols facilitate applications in research, medicine, and agriculture.