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High efficiency gene transfer into mammalian cells.

C M Gorman, D P Lane, P W Rigby

    Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
    |December 4, 1984
    PubMed
    Summary

    This study presents a generalized gene transfer protocol, expanding recipient cell types for foreign gene insertion. The optimized transient assay system enables sensitive detection of gene function in various cells, including human and embryonic stem cells.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Cell Biology

    Background:

    • Gene transfer is crucial for genetic research and therapeutic applications.
    • Existing methods often have limitations in recipient cell type and efficiency.
    • Developing versatile gene transfer protocols is essential for advancing biological studies.

    Purpose of the Study:

    • To generalize and optimize a gene transfer protocol for a wider range of recipient cells.
    • To establish a rapid screening system for foreign DNA expression.
    • To enable sensitive detection of gene function in stable transformants.

    Main Methods:

    • Utilized a transient assay system for rapid screening of foreign DNA expression.
    • Optimized gene transfer conditions using the transient system.

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  • Applied optimized conditions for efficient stable transformation.
  • Tested the protocol on primate, human, and undifferentiated embryonic cells.

    • Main Results:

    • Successfully generalized the gene transfer protocol to a broader spectrum of cells.
    • The transient assay system allowed for efficient screening and optimization.
    • Achieved sensitive detection of single-copy gene function in human cells.
    • Demonstrated successful gene transfer in undifferentiated embryonic cells.

    Conclusions:

    • The generalized gene transfer protocol significantly expands the utility of gene transfer techniques.
    • The transient assay system is effective for optimizing gene transfer efficiency.
    • This method facilitates sensitive genetic analysis in various cell types, including human and stem cells.
    • The protocol holds promise for applications in genetic research and biotechnology.