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Cloning Mice

Atsuo Ogura

    Cold Spring Harbor Protocols
    |August 3, 2017
    PubMed

    Insights

    Somatic nuclear transfer (SCNT) enables the creation of viable cloned mice by reprogramming donor cell DNA within oocytes. Efficient micromanipulation techniques improve success rates in this mouse cloning method.

    Area of Science:

    • Reproductive Biology
    • Developmental Biology
    • Genetics

    Background:

    • Somatic nuclear transfer (SCNT) is a technique used to generate cloned offspring.
    • Reprogramming of the donor cell genome within the oocyte is crucial for successful SCNT.
    • Previous methods for mouse cloning involved direct injection of donor nuclei into oocytes.

    Purpose of the Study:

    • To describe a refined protocol for somatic nuclear transfer in mice.
    • To highlight key factors contributing to the success of mouse cloning via SCNT.
    • To detail the steps involved in generating cloned mice using the Honolulu method.

    Main Methods:

    • Somatic nuclear transfer (SCNT) using enucleated oocytes and donor nuclei.
    • Utilizing blunt micropipettes with a piezo impact-driving micromanipulation device for efficiency.
    • A seven-part protocol including oocyte preparation, nuclear injection, embryo activation, and transfer.

    Main Results:

    • Viable and fertile cloned mice were generated through SCNT.
    • Donor nuclei readily condensed within oocytes, facilitating genome reprogramming.
    • The use of advanced micromanipulation tools significantly improved oocyte viability and procedural efficiency.

    Conclusions:

    • SCNT is an effective method for producing cloned mice.
    • Oocyte factors and efficient micromanipulation are critical for successful mouse cloning.
    • The described protocol provides a robust approach to SCNT in mice.

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