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Conservative Site-specific Recombination and Phase Variation02:53

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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
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Binary recombinase systems for high-resolution conditional mutagenesis.

Mario Hermann1, Patrick Stillhard, Hendrik Wildner

  • 1Institute of Laboratory Animal Science, University of Zurich, Sternwartstrasse 6, CH-8091 Zurich, Switzerland, Institute of Neuropathology, University Hospital of Zurich, Schmelzbergstrasse 12, CH-8091 Zurich, Switzerland, Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland, National Centre for Biotechnology (CNB-CSIC), Darwin 3, 28049 Madrid, Spain, CIBERER-ISCIII, Darwin 3, 28049 Madrid, Spain, Program of Cardiovascular Development, Department of Cardiovascular Development and Repair, Centro Nacional de Investigaciones Cardiovasculares Carlos III, calle Melchor Fernández Almagro 3, 28029 Madrid, Spain and Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.

Nucleic Acids Research
|January 14, 2014
PubMed
Summary
This summary is machine-generated.

Two new systems, Co-Driver and Co-InCre, enhance conditional mutagenesis for precise gene function studies and lineage tracing. These tools improve recombination efficiency and accuracy, expanding the capabilities of Cre-lox technology in biological research.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • Conditional mutagenesis using Cre recombinase is crucial for gene function and lineage tracing.
  • Existing systems may lack accuracy or optimal recombination efficiency.

Purpose of the Study:

  • To introduce two novel dual-promoter-driven conditional mutagenesis systems: Co-Driver and Co-InCre.
  • To enhance accuracy and efficiency of recombination for genetic analyses.

Main Methods:

  • Co-Driver utilizes a Dre-Cre recombinase cascade for sequential recombination.
  • Co-InCre employs trans-splicing inteins and simultaneous promoter activity to reconstitute Cre recombinase.

Main Results:

  • Co-Driver enables precise sequential lineage tracing by requiring a specific temporal order of recombinase expression.
  • Co-InCre achieves high recombination rates by generating native Cre recombinase, outperforming other binary systems.
  • Both systems demonstrate significant improvements in the utility of existing Cre-responsive alleles.

Conclusions:

  • Co-Driver and Co-InCre represent significant advancements in conditional mutagenesis technologies.
  • These systems offer greater accuracy and efficiency for gene function studies and lineage tracing.