When eggs and cleaving embryos of the sea urchin are exposed to [(3)H]actinomycin D, they become radioactive, and autoradiograms show that the radioactivity is inside the cells. At midcleavage, nuclei are more radioactive than cytoplasm. Extraction and chromatography of the intracellular labeled compounds identify them as actinomycin D and a water-soluable derivative. Conversion does not take place outside the cells. Treatment of embryos for 90 minutes with actinomycin D inhibits synthesis of RNA by more than 90 percent, leaving unaffected turnover of the pCpCpA terminals in transfer RNA. These data justify earlier interpretations of actinomycin experiments with embryos and justify use of the drug as a tool in the analysis of gene expression.