MeCP2 CRISPR Activation Plasmid (h): sc-401228-ACT

MECP2 encodes methyl-CpG binding protein 2 (MeCP2), a chromatin-associated regulator that binds methylated DNA and shapes transcriptional programs by coordinating epigenetic repression and activation. MeCP2 influences neuronal maturation, synaptic plasticity, and activity-dependent gene expression through interactions with corepressors and chromatin remodeling complexes, linking DNA methylation dynamics to RNA processing and genome stability. Dysregulated MECP2 dosage perturbs neurodevelopmental pathways and alters excitatory–inhibitory balance, making it highly relevant for studies of transcriptional control in differentiated neurons and glia. Because MeCP2 integrates epigenetic signaling with long-range chromatin architecture, it is widely used as a model to interrogate gene regulatory networks in the human nervous system.

MeCP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MECP2 expression without altering the underlying DNA sequence.

MeCP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MECP2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the MECP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MeCP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MECP2 locus and enabling the study of MeCP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MeCP2 pathway restoration in tumor cells with silenced or reduced MECP2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.