SAF-B CRISPR Activation Plasmid (h): sc-403795-ACT

SAFB (SAF-B, scaffold attachment factor B) is a nuclear matrix–associated DNA- and RNA-binding protein that coordinates chromatin organization with transcriptional control and pre-mRNA processing. It interacts with scaffold/matrix attachment regions and regulates gene expression programs linked to RNA polymerase II activity, alternative splicing, and stress-responsive transcription. SAF-B has been implicated in hormone receptor signaling, particularly estrogen receptor–associated regulatory networks, and contributes to maintaining genome function through coupling of nuclear architecture to RNA metabolism. Dysregulation of SAFB expression or function has been associated with altered transcriptional states observed in cancer-related pathways and other disorders involving aberrant gene regulation.

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

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

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