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Related Concept Videos

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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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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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Related Experiment Video

Updated: Dec 16, 2025

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain
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Neural circuit analysis using a novel intersectional split intein-mediated split-Cre recombinase system.

Audrey Tze Ting Khoo1, Paul Jong Kim1, Ho Min Kim2,3

  • 1Neuroscience and Behavioural Disorders Programme, Duke-National University of Singapore (NUS) Medical School, 8 College Road, Singapore, 169857, Singapore.

Molecular Brain
|July 4, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel split Cre recombinase system to precisely label specific neuron types. This method combines gene expression and connectivity for advanced brain circuit analysis.

Keywords:
Circuit mappingDLX6GABAProjection neuronsRetrograde virusSplit inteinSplit-cre recombinaseTransgenic mouse

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Neurons form complex networks essential for animal behavior.
  • Current methods for targeting neuronal populations have limitations in specificity.
  • Understanding neural circuits requires precise tools to delineate specific neuron types.

Purpose of the Study:

  • To develop a novel system for selective labeling of specific neuronal populations.
  • To overcome limitations of existing methods in distinguishing neuronal types based on gene expression and connectivity.
  • To enable sophisticated characterization of mammalian brain circuits.

Main Methods:

  • Developed an intersectional split intein-mediated split-Cre recombinase system.
  • Split Cre recombinase into two fragments using evolved split inteins.
  • Expressed one fragment via a cell type-specific promoter in transgenic animals.
  • Delivered the second fragment using retrograde viral gene transfer.

Main Results:

  • Achieved reconstitution of Cre recombinase activity specifically in targeted neuronal populations.
  • Enabled selective labeling based on both gene expression profiles and structural connectivity.
  • Demonstrated the system's ability to target neurons projecting from specific brain regions or of specific neuronal types.

Conclusions:

  • The novel split intein-based split-Cre system offers high specificity for neuronal population labeling.
  • This system provides a powerful tool for advanced characterization of mammalian neural circuits.
  • Facilitates detailed investigation into the function of specific neuronal types within complex brain networks.