Gene expression regulation is an important molecular process for monitoring overall expression level of genes in plants. The expression of genes is regulated by certain regulatory elements including, Cis-acting regulatory elements (CRE), transcription factors (TFs) and promoters. Cis-acting elements are actually specific class of DNA binding proteins that act at particular site of DNA. Promoters are part of DNA fragment essential for transcriptional regulation of genes with certain transcription factors. These transcription factors could be useful in developmental regulation, interpretation and validation of candidate genes. This review highlights the importance of Cis-acting regulatory elements and transcription factors for regulation of gene expression. Furthermore, the use of bioinformatics approach for identification of transcription factors and putative motifs within the promoter region has gained much success for studying DNA-protein interactions. These findings promote the importance of CRE and TF inactivation of transcriptional networks for enhanced gene expression studies in plants.