NACA1 CRISPR Activation Plasmid (h): sc-408139-ACT

Human NACA encodes nascent polypeptide–associated complex subunit alpha (NACA1), a ribosome-associated chaperone that binds emerging polypeptide chains and helps coordinate co‑translational folding, targeting, and quality control. NACA1 functions with its partner subunit to modulate nascent chain interactions at the ribosomal exit tunnel, influencing protein biogenesis and proteostasis pathways linked to endoplasmic reticulum targeting and stress-adaptive responses. By shaping the balance between productive translation and misfolded protein handling, NACA1 is relevant to cellular states characterized by altered translational control, including proliferative programs and stress signaling. Dysregulation of NACA/NACA1 expression has been reported across multiple disease-associated contexts, motivating mechanistic studies of how co‑translational regulation impacts cell fate and transcriptional networks.

NACA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NACA expression without altering the underlying DNA sequence.

NACA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NACA 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 NACA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NACA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NACA locus and enabling the study of NACA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NACA1 pathway restoration in tumor cells with silenced or reduced NACA expression.

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