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Cellular and molecular studies on globin gene expression

W F Anderson, A Deisseroth, A W Nienhuis

    National Cancer Institute Monograph
    |May 1, 1978
    PubMed
    Summary
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    Somatic cell hybrids reveal that globin gene expression is regulated by cytoplasmic factors, not nuclear ones. Microinjection into Chinese hamster ovary cells demonstrates a method for assaying these regulatory molecules.

    Area of Science:

    • Cell Biology
    • Molecular Biology
    • Genetics

    Background:

    • Globin gene expression is crucial for hemoglobin production.
    • Previous studies utilized somatic cell hybrids to investigate gene regulation.

    Purpose of the Study:

    • To investigate the regulatory mechanisms controlling globin gene expression.
    • To develop an intracellular assay for identifying globin gene regulatory molecules.

    Main Methods:

    • Somatic cell hybridization between mouse erythroleukemia (MEL) and human erythroid/non-erythroid cells.
    • Molecular hybridization techniques (DNA-RNA) to detect globin genes and mRNA.
    • Cybrid formation using enucleated nonerythroid cells.
    • Microinjection of rabbit globin mRNA into Chinese hamster ovary (CHO) cells using red blood cell (RBC) ghosts.

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    Main Results:

    • MEL-human erythroid hybrids coexpressed human and mouse globin genes.
    • MEL-non-erythroid hybrids lost globin gene expression, indicating post-transcriptional or transcriptional regulation.
    • Cybrids confirmed that the nucleus is not required for globin gene silencing.
    • Microinjected CHO cells synthesized hemoglobin, validating the microinjection technique.

    Conclusions:

    • Globin gene expression is regulated by diffusible cytoplasmic factors.
    • The developed microinjection technique offers a potential assay for intracellular regulatory molecules.
    • Cellular cytoplasm plays a critical role in controlling gene expression patterns.