WSTF CRISPR Activation Plasmid (h): sc-404237-ACT

Human BAZ1B encodes Williams syndrome transcription factor (WSTF), a nuclear chromatin-associated protein that integrates ATP-dependent remodeling with histone H2A.X phosphorylation during DNA damage responses. WSTF functions in the WICH complex with SMARCA5 and also contributes to ISWI-family chromatin remodeling, linking nucleosome positioning to transcriptional regulation and replication-associated chromatin maintenance. Through these activities, BAZ1B influences genome stability, cell-cycle progression, and RNA polymerase II–dependent gene expression programs. Altered BAZ1B dosage and dysregulated WSTF-dependent chromatin states have been associated with neurodevelopmental phenotypes and broader transcriptional perturbations relevant to disease biology.

WSTF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BAZ1B expression without altering the underlying DNA sequence.

WSTF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BAZ1B 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 BAZ1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous WSTF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BAZ1B locus and enabling the study of WSTF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of WSTF pathway restoration in tumor cells with silenced or reduced BAZ1B expression.

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