SPT16 CRISPR Activation Plasmid (h): sc-401268-ACT

Human SUPT16H encodes SPT16, a core subunit of the FACT (Facilitates Chromatin Transcription) histone chaperone complex that reorganizes nucleosomes to support RNA polymerase II elongation. By coordinating H2A–H2B dynamics and chromatin accessibility, SPT16 helps regulate transcriptional programs, replication-associated chromatin remodeling, and DNA damage responses. FACT-dependent chromatin control links SUPT16H to proliferative signaling networks and genome stability pathways, making it relevant for studying epigenetic regulation and mechanisms that contribute to oncogenic transcription and stress adaptation. Altered FACT activity has been associated with dysregulated gene expression and chromatin states observed across multiple cancer-related contexts, supporting its use as a functional node in chromatin and transcription research.

SPT16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SUPT16H expression without altering the underlying DNA sequence.

SPT16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SUPT16H 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 SUPT16H transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPT16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SUPT16H locus and enabling the study of SPT16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPT16 pathway restoration in tumor cells with silenced or reduced SUPT16H expression.

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