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Related Experiment Videos

Conditional gene knockout using Cre recombinase.

Y Le, B Sauer

    Molecular Biotechnology
    |July 4, 2001
    PubMed
    Summary
    This summary is machine-generated.

    Cre recombinase enables precise genetic manipulation in mice by excising DNA sequences flanked by loxP sites. Reliable detection of Cre gene expression is crucial for successful conditional mutagenesis, using methods like PCR and GFP-tagged Cre.

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    Area of Science:

    • Genetics and Genomics
    • Molecular Biology
    • Developmental Biology

    Background:

    • Cre recombinase is a key tool for genetic engineering in mice.
    • Conditional mutagenesis relies on precise DNA sequence excision mediated by Cre recombinase and loxP sites.
    • Effective conditional mutagenesis requires careful allele design and reliable Cre gene expression detection.

    Purpose of the Study:

    • To highlight the importance of Cre recombinase in genetic manipulation.
    • To emphasize the necessity of accurate Cre gene expression detection for conditional mutagenesis.
    • To outline methods for verifying Cre activity in genetically modified mice.

    Main Methods:

    • Polymerase Chain Reaction (PCR) for DNA analysis.
    • Immunohistochemistry for protein localization.

    Related Experiment Videos

  • Utilizing a recombination-proficient Green Fluorescent Protein (GFP)-tagged Cre protein for detection.

    • Main Results:

    • Demonstrated the utility of Cre-loxP system for targeted DNA excision.
    • Validated various methods for detecting Cre expression and recombination.
    • Showcased the importance of reliable Cre detection for experimental success.

    Conclusions:

    • Cre recombinase is indispensable for conditional mutagenesis in mice.
    • Accurate detection of Cre gene expression is critical for successful genetic manipulation.
    • A combination of molecular and protein-based methods ensures reliable verification of Cre activity.