The biological effect of RNA preparations from the thyroid glands of TSH-stimulated guinea pigs was studied. In particular, iodine metabolism of the thyroid gland was stimulated by both TSH and thyroid RNA in vitro but not by liver RNA or RNase-treated thyroid RNA. The results indicate that thyroid RNA mediates the TSH stimulated iodine metabolism in the thyroid. This TSH-like effect of thyroid RNA is organ-specific and not due to contamination of RNA with TSH.