TUSC3 CRISPR Activation Plasmid (h): sc-405571-ACT

TUSC3 (tumor suppressor candidate 3) encodes an endoplasmic reticulum membrane protein that functions as a subunit of the oligosaccharyltransferase (OST) complex, supporting co-translational N-linked glycosylation and quality control of nascent secretory and membrane proteins. Through its role in protein folding homeostasis, TUSC3 influences ER proteostasis, glycoprotein maturation, and downstream signaling processes sensitive to glycosylation status, including receptor trafficking and stress-adaptive responses. Altered TUSC3 expression or loss-of-function has been linked to neurodevelopmental phenotypes and has been studied in oncology contexts where disrupted glycosylation and ER stress pathways can reshape proliferation, adhesion, and migration programs. These features make TUSC3 a useful node for investigating glycoproteostasis, ER-associated pathways, and gene-regulatory networks relevant to disease biology in human cell models.

TUSC3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TUSC3 expression without altering the underlying DNA sequence.

TUSC3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TUSC3 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 TUSC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TUSC3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TUSC3 locus and enabling the study of TUSC3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TUSC3 pathway restoration in tumor cells with silenced or reduced TUSC3 expression.

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