REST-VP16 activates multiple neuronal differentiation genes in human NT2 cells

The RE1-silencing transcription factor (REST)/neuron-restrictive silencer f actor (NRSF) can repress transcription of a battery of neuronal differentia tion genes in non-neuronal cells by binding to a specific consensus DNA seq uence present in their regulatory regions. However, REST/NRSF-/- mice sugge st that the absence of REST/NRSF-dependent repression alone is not sufficie nt for the expression of these neuronal differentiation genes and that the presence of other promoter/enhancer-specific activators is required. Here w e describe the construction of a recombinant transcription factor, REST-VP1 6, by replacing repressor domains of REST/NRSF with the activation domain o f a viral activator VP16, In transient transfection experiments, REST-VP16 was found to operate through RE1 binding site/neuron-restrictive enhancer e lement (RE1/NRSE), activate plasmid-encoded neuronal promoters in various m ammalian cell types and activate cellular REST/ NRSF target genes, even in the absence of factors that are otherwise required to activate such genes. Efficient expression of REST-VP16 through adenoviral vectors in NT2 cells, which resemble human committed neuronal progenitor cells, was found to caus e activation of multiple neuronal genes that are characteristic markers for neuronal differentiation. Thus, REST-VP16 could be used as a unique tool t o study neuronal differentiation pathways and neuronal diseases that arise due to the deregulation of this process.