Small RNAs (sRNAs) that act as regulators of gene expression have been identified in all kingdoms of life. Until 1999, only about 10 abundant sRNAs had been identified in Escherichia coli, but the function of most of them remained elusive for a long time. However, since 2001, a series of systematic computational approaches have revealed that bacteria encode a tremendous number of sRNAs. In E. coli more than 100 sRNAs are now known. However, approximately only 20 of them have been assigned a biological function, indicating that this is still a challenging issue. Systematic searches have been performed for a few Gram-positive bacterial species, too. sRNAs can be divided into two major groups: the first group comprises so-called bona fide antisense RNAs, which regulate gene expression by a base-pairing mechanism with mRNA. The second group of sRNAs encompasses RNAs that act by binding to small proteins.