CHD7 CRISPR Activation Plasmid (h): sc-404017-ACT

CHD7 (chromodomain helicase DNA binding protein 7) is an ATP-dependent chromatin remodeler that interprets histone marks via its chromodomains to regulate enhancer and promoter accessibility. In human cells, CHD7 coordinates transcriptional programs that shape lineage specification, neural crest development, and organogenesis by modulating nucleosome positioning and higher-order chromatin architecture. CHD7 function intersects with core epigenetic and transcriptional regulatory pathways, influencing RNA polymerase II–driven gene expression and chromatin state transitions during differentiation. Genetic disruption or altered dosage of CHD7 is linked to developmental disorder phenotypes, including CHARGE syndrome, and its dysregulation is investigated in neurodevelopmental and cancer-associated transcriptional networks.

CHD7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHD7 expression without altering the underlying DNA sequence.

CHD7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHD7 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 CHD7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHD7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHD7 locus and enabling the study of CHD7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHD7 pathway restoration in tumor cells with silenced or reduced CHD7 expression.

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