Synthetic Promoter Elem Sequence Variation and S

Eukaryotic transcriptional regulation in different cells invc large numbers and arrangements of cis and trans elements survey the number of cis regulatory elements that are activ different contexts, we have devised a high-throughput selec procedure permitting synthesis of active cis motifs that enh the activity of a minimal promoter. This synthetic promoter struction method (SPCM) was used to identify >100 DNA quences that showed increased promoter activity in the ne blastoma cell line Neuro2A. After determining DNA sequence selected synthetic promoters, database searches for known ments revealed a predominance of eight motifs: AP2, CEBP, I Ebox, ETS, CREB, AP1, and SP1/MAZ. The most active of selected synthetic promoters contain composites of a numbE these motifs. Assays of DNA binding and promoter activity of t exemplary motifs (ETS, CREB, and SP1/MAZ) were used to p the effectiveness of SPCM in uncovering active sequences. U 10% of 133 selected active sequences had no match in curre available databases, raising the possibility that new motifs transcriptional regulatory proteins to which they bind ma! revealed by SPCM. The method may find uses in construe databases of active cis motifs, in diagnostics, and in gene ther