Sec61α1 CRISPR Activation Plasmid (h): sc-402222-ACT

SEC61A1 encodes the alpha subunit of the Sec61 translocon, a conserved protein-conducting channel in the endoplasmic reticulum membrane that mediates co-translational insertion and translocation of nascent polypeptides. Sec61α1 functions at the interface of ribosome docking, signal peptide recognition, and ER protein quality control, linking secretory pathway biogenesis to proteostasis and the unfolded protein response. By governing ER entry of secreted and membrane proteins, SEC61A1 influences processes such as glycoprotein maturation, ER-associated degradation, and cellular stress signaling. Perturbation of ER translocation capacity is relevant to disorders driven by proteostasis imbalance and secretory pathway dysfunction, supporting its study in cell survival, differentiation, and stress-adaptation contexts.

Sec61α1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEC61A1 expression without altering the underlying DNA sequence.

Sec61α1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEC61A1 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 SEC61A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sec61α1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEC61A1 locus and enabling the study of Sec61α1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sec61α1 pathway restoration in tumor cells with silenced or reduced SEC61A1 expression.

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