SPT6 CRISPR Activation Plasmid (h): sc-406219-ACT

Human SUPT6H encodes SPT6, a conserved transcription elongation and chromatin-associated factor that binds the phosphorylated RNA polymerase II CTD and coordinates nucleosome reassembly during active transcription. SPT6 functions with factors such as IWS1/ALYREF to couple elongation with co‑transcriptional mRNA processing, including splicing and RNA export, thereby shaping transcriptional fidelity and genome stability. Through regulation of chromatin structure and elongation dynamics, SUPT6H influences broad gene expression programs linked to cell identity, proliferation, and stress responses. Dysregulated SPT6-dependent transcriptional control has been implicated in aberrant oncogenic signaling and altered differentiation states, making it a useful node for mechanistic studies of transcriptional addiction and epigenetic vulnerability.

SPT6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SUPT6H expression without altering the underlying DNA sequence.

SPT6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SUPT6H 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 SUPT6H transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPT6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SUPT6H locus and enabling the study of SPT6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPT6 pathway restoration in tumor cells with silenced or reduced SUPT6H expression.

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