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Related Concept Videos

Methods of Nuclear Reprogramming01:24

Methods of Nuclear Reprogramming

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Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for...
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Nuclear reprogramming is the process of switching gene expression of one cell type to that of another cell type, usually from a differentiated cell state to an undifferentiated cell state. Differentiation occurs during processes such as development and morphogenesis, tissue regeneration, and malignancy. Cells can also be artificially induced to reprogram their gene expression by techniques such as nuclear transfer, induced pluripotency, and cell fusion. Such techniques have many applications in...
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Related Experiment Video

Updated: Apr 1, 2026

Transnuclear Mice with Pre-defined T Cell Receptor Specificities Against Toxoplasma gondii Obtained Via SCNT
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Generation of Thy1 Constructs for Pronuclear Injection.

Petar Marinković, Leanne Godinho, Thomas Misgeld

    Cold Spring Harbor Protocols
    |October 3, 2015
    PubMed
    Summary

    Generating Thy1-XFP transgenic mice is now accessible for small labs. This protocol details cloning fluorescent reporter proteins into the Thy1 vector and preparing DNA for pronuclear injection.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Neuroscience

    Background:

    • Generating transgenic mouse models is crucial for studying gene function.
    • Thy1 promoter-driven expression is widely used for neuronal targeting.
    • Small laboratories face challenges in generating custom transgenic lines.

    Purpose of the Study:

    • To provide a detailed protocol for key steps in generating Thy1-XFP transgenic mice.
    • To simplify the process of cloning fluorescent reporter proteins into the Thy1 vector.
    • To enable small laboratories to create their own transgenic lines.

    Main Methods:

    • Cloning of fluorescent reporter protein sequences into the XhoI site of the Thy1 vector using blunt-end cloning.
    • Linearization of the Thy1 construct using EcoRI and PvuI restriction enzymes.

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  • Purification of the linearized DNA for pronuclear injection.

    • Main Results:

    • A reliable method for cloning reporter genes into the Thy1 vector is presented.
    • The protocol ensures proper preparation of DNA constructs for pronuclear injection.
    • Successful generation of founder mice is dependent on specialized injection facilities.

    Conclusions:

    • This protocol facilitates the generation of Thy1-XFP transgenic mice, even in resource-limited settings.
    • The described cloning and linearization steps are critical for successful transgenic line creation.
    • Accessible generation of transgenic models accelerates neuroscience research.