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Integration-specific retrovirus expression in embryonal carcinoma cells.
Summary
Undifferentiated embryonal carcinoma (EC) cells, typically resistant to gene expression, showed transcription of a small fraction of retroviruses. This suggests a cis-acting element or viral mutations may enable gene expression in these stem cells.
Area of Science:
- * Molecular Biology
- * Developmental Biology
- * Virology
Background:
- * Embryonal carcinoma (EC) cells are pluripotent stem cells.
- * EC cells are generally resistant to retroviral gene expression.
Purpose of the Study:
- * To investigate retroviral gene expression in undifferentiated EC cells.
- * To identify factors influencing provirus transcription in EC cells.
Main Methods:
- * Infection of PCC4 aza 1 EC cells with a selectable amphotropic retrovirus.
- * Cloning and expansion of cells with active proviruses.
- * Nucleic acid analysis to quantify proviral RNA levels and assess gene expression.
Main Results:
- * Approximately 1 in 5,000-10,000 proviruses were transcribed in EC cells.
- * Viral RNA levels varied among cell clones (0.05%-0.5% of poly(A)+ RNA).
- * EC cells maintained an undifferentiated phenotype and blocked expression of other integrated proviruses.
Conclusions:
- * Retroviral expression in EC cells is rare but achievable.
- * Cis-acting genetic elements at integration sites or viral mutations may mediate expression.
- * Findings provide insights into gene regulation in pluripotent stem cells.