SGTA CRISPR Activation Plasmid (h): sc-404399-ACT

Human SGTA (small glutamine-rich tetratricopeptide repeat-containing protein alpha) is a co-chaperone that binds HSP70/HSC70 and HSP90 to coordinate protein quality control and client protein handling. SGTA participates in triage of hydrophobic polypeptides and tail-anchored membrane proteins, influencing their delivery to membrane insertion pathways or proteasomal degradation, and it interfaces functionally with the BAG6/GET targeting axis. Through these interactions, SGTA helps maintain proteostasis, modulates receptor and signaling protein maturation, and impacts cellular stress responses such as ER-associated degradation (ERAD). Dysregulation of chaperone networks that include SGTA has been associated with altered protein homeostasis in contexts relevant to cancer biology and neurodegeneration, supporting its value as a mechanistic node for pathway interrogation.

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

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

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