SUGT1 CRISPR Activation Plasmid (h): sc-405128-ACT

SUGT1 (SGT1 homolog) encodes a conserved co-chaperone that couples HSP90 activity to the assembly and stability of multiprotein complexes, including kinetochore components required for accurate chromosome segregation. In human cells, SUGT1 supports cell-cycle progression through mitotic checkpoint control and contributes to ubiquitin–proteasome-related quality control by coordinating protein complex maturation. Through these roles, SUGT1 is relevant to pathways governing genome stability, aneuploidy, and stress-responsive signaling networks. Dysregulated SUGT1 function or expression has been investigated in contexts where proliferative control and proteostasis are altered, providing a mechanistic entry point for studying cell division defects and cancer-associated phenotypes.

SUGT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SUGT1 expression without altering the underlying DNA sequence.

SUGT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SUGT1 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 SUGT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SUGT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SUGT1 locus and enabling the study of SUGT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SUGT1 pathway restoration in tumor cells with silenced or reduced SUGT1 expression.

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