orientation-dependent but must also use a promoter-specific factor. There has been a recent discovery of Polycomb-group Response Elements (PREs), which can allow and inhibit repression depending on the protein bound to it, and the presence of non-coding transcription. Mechanisms For classical silencers, the signaling pathway is relatively simple. Since repression is active, silencer elements target the assembly of GTFs, necessary for transcription of the gene. These silencer elements are mostly located upstream of the gene and can vary between short and long distances. For long-range silencers, it has been observed that the DNA will form a loop in order to bring the silencer closer to the promoter and loop out the interfering DNA. Silencers also target helicase sites in the DNA that are rich in adenine and thymine (AT) and prone to unwinding the DNA, allowing room to initiate transcription. The inhibited helicase activity leads to the inhibition of transcription. This is commonly seen in the human thyrotropin-β gene promoter. NREs can induce a bend in the promoter region to block interactions, as seen when an NRE binds to Yin-Yang 1 (YY1), and flank regulatory signals or promoter regions as well. When the silencer region is located within an intron, there can be two types of repressions. First, there can be a physical blockage of a splice site. Second, there can be a bend in the DNA that will inhibit RNA processing. When located in the exon or the untranslated region, the silencer will mainly be classical or position-dependent. However, these silencers can carry out their activity prior to transcription. Most silencers are constitutively expressed in organisms, only allowing activation of a gene by either inhibiting the silencer or by activating an enhancer region. The best example of this is the Neuronal-Restrictive Silencer Factor (NRSF) that is produced by the REST gene. The REST gene produces NRSF in order to repress the transcription of neuronal genes that are essential for localization of neuronal tissue. When a silencer represses REST, NRSF is also inhibited, allowing for the transcription of neuronal genes. Similarities with enhancers Another regulatory element located upstream of the gene is an enhancer. Enhancers function as a "turn on" switch in gene expression and will activate the promoter region of a particular gene while silencers act as the "turn off" switch. Though these two regulatory elements work against each other, both sequence types affect the promoter region in very