Set1A CRISPR Activation Plasmid (h): sc-406841-ACT

Human SETD1A encodes Set1A, a catalytic component of the COMPASS histone methyltransferase complex that deposits H3K4 methylation marks associated with transcriptionally active chromatin. Through regulation of promoter-proximal chromatin states, Set1A influences RNA polymerase II–dependent transcription, enhancer–promoter communication, and cell state maintenance programs during development and differentiation. SETD1A function is linked to genome integrity and replication-associated processes via coordination of chromatin accessibility and DNA damage response signaling. Dysregulation of SETD1A-mediated H3K4 methylation has been associated with neurodevelopmental and neuropsychiatric phenotypes and altered transcriptional networks relevant to human disease biology.

Set1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SETD1A expression without altering the underlying DNA sequence.

Set1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SETD1A 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 SETD1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Set1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SETD1A locus and enabling the study of Set1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Set1A pathway restoration in tumor cells with silenced or reduced SETD1A expression.

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