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Related Concept Videos

Plasmids01:28

Plasmids

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Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...
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Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Related Experiment Video

Updated: Mar 17, 2026

High-Throughput DNA Plasmid Multiplexing and Transfection Using Acoustic Nanodispensing Technology
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Evidence for plasmid DNA exchange after polyplex mixing.

L Pigeon1, C Gonçalves1, C Pichon1

  • 1Centre de Biophysique Moléculaire, CNRS UPR4301, Inserm and University of Orléans, 45071 Orléans cedex 02, France. patrick.midoux@cnrs-orleans.fr.

Soft Matter
|July 28, 2016
PubMed
Summary
This summary is machine-generated.

When two different fluorescently labeled plasmid DNA (pDNA) polyplexes are mixed, pDNA exchange occurs, indicating polyplex unpackaging and reassembly. This exchange is dependent on the polymer type and pDNA interaction strength.

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Area of Science:

  • Biochemistry
  • Materials Science
  • Nanotechnology

Background:

  • Cationic polymers and liposomes self-assemble with plasmid DNA (pDNA) to form nanostructures like polyplexes and lipoplexes.
  • These nanostructures are crucial for gene delivery applications.
  • Understanding their stability and dynamics is key to optimizing delivery efficiency.

Purpose of the Study:

  • To investigate the exchange of plasmid DNA (pDNA) between pre-formed polyplexes.
  • To determine the factors influencing pDNA exchange dynamics.
  • To elucidate the mechanism of pDNA exchange in polyplexes.

Main Methods:

  • Preparation of two distinct fluorescently labeled pDNA/histidinylated lPEI polyplexes (Flu-pDNA/His-lPEI and Cy5-pDNA/His-lPEI).
  • Mixing of the two polyplex preparations.
  • Analysis of dual fluorescence and Förster Resonance Energy Transfer (FRET) to detect pDNA exchange.
  • Comparison with other polyplexes (polylysine, caged His-lPEI) and other nanostructures (lipoplexes, lipopolyplexes).

Main Results:

  • Mixing Flu-pDNA/His-lPEI and Cy5-pDNA/His-lPEI polyplexes resulted in a high proportion of dual-fluorescent polyplexes.
  • FRET analysis confirmed the presence of both types of fluorescent pDNA within single polyplexes, indicating successful pDNA exchange.
  • pDNA exchange was observed to be dependent on the polymer type and the strength of pDNA/polymer interactions.
  • No pDNA exchange was detected with polylysine polyplexes, caged His-lPEI polyplexes, lipoplexes, lipopolyplexes, or when His-lPEI polyplexes were mixed with lipoplexes.

Conclusions:

  • Polyplexes formed with histidinylated lPEI facilitate pDNA exchange between distinct polyplexes.
  • The observed pDNA exchange suggests a mechanism involving polyplex aggregation, unpackaging, and subsequent reassembly.
  • The findings highlight the dynamic nature of certain polyplex structures and are critical for understanding gene delivery vector stability.