Set1B CRISPR Activation Plasmid (h): sc-406129-ACT

Human SETD1B encodes Set1B, a SET domain–containing histone methyltransferase that catalyzes H3K4 methylation associated with transcriptionally active chromatin. As a component of COMPASS-like complexes, Set1B helps coordinate promoter-proximal chromatin states, transcription initiation, and enhancer–promoter communication, integrating with broader epigenetic regulation and RNA polymerase II–dependent gene expression programs. Perturbation of SETD1B function is linked to dysregulated transcriptional control and altered developmental gene networks, with relevance to neurodevelopmental phenotypes and other contexts where chromatin-dependent gene regulation is disrupted. These features make SETD1B a useful locus for interrogating how H3K4 methylation shapes lineage specification, neuronal function, and stimulus-responsive transcription.

Set1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SETD1B expression without altering the underlying DNA sequence.

Set1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SETD1B 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 SETD1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Set1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SETD1B locus and enabling the study of Set1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Set1B pathway restoration in tumor cells with silenced or reduced SETD1B expression.

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