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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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.
The recognition sites for Cre recombinase called LoxP...
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Gene Conversion02:08

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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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Restriction Enzymes01:11

Restriction Enzymes

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Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
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Homologous Recombination02:31

Homologous Recombination

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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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Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

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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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Combinatorial Gene Control02:33

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Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
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Related Experiment Video

Updated: May 9, 2025

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
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Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)

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[Construction of an intein-mediated Split-Cre system].

Yifei Ao1, Qi Zhang2, Yuxi Chen3

  • 1School of Life Sciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, Guangdong, China.

Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology
|May 6, 2025
PubMed
Summary

Researchers developed an efficient Split-Cre system using Rma intein. This novel Split-Cre system demonstrated comparable efficiency to full-length Cre in mice.

Keywords:
CreRhodothermus marinus (Rma) inteinSplit-CreSplit-intein

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Last Updated: May 9, 2025

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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Area of Science:

  • Molecular Biology
  • Biotechnology

Context:

  • The Split-Cre system, comprising NCre and CCre, enables Cre recombinase activity upon reassembly.
  • Cre-loxP recombination is a fundamental tool in genetic engineering and research.

Purpose:

  • To engineer an efficient Split-Cre system using intein-mediated protein splicing.
  • To identify an optimal split site for Cre recombinase activity.

Summary:

  • This study utilized Rma intein from Rhodothermus marinus to split Cre recombinase.
  • The split site S102 was identified for efficient Cre-mediated recombination in a reporter cell line.
  • The S102 Split-Cre system, delivered via dual adeno-associated virus (AAV) vectors in mice, showed recombination efficiency comparable to full-length Cre.

Impact:

  • Establishes a functional Split-Cre system for advanced genetic manipulation.
  • Provides a foundation for diverse applications in basic and applied biological research.