HSPA2 CRISPR Activation Plasmid (h): sc-400832-ACT

HSPA2 encodes a member of the HSP70 family of ATP-dependent molecular chaperones that supports proteostasis by promoting nascent polypeptide folding, preventing protein aggregation, and coordinating refolding or turnover of stress-damaged proteins. HSPA2 activity interfaces with heat shock response regulation, chaperone-mediated protein quality control, and cellular adaptation to proteotoxic stress, influencing processes such as cell cycle progression and differentiation. In human tissues, HSPA2 is notably linked to germ cell development and sperm function, and altered expression has been reported across disease-relevant contexts where proteostasis and stress signaling are perturbed, including tumor biology and neurodegeneration. These features make HSPA2 a useful node for investigating stress-responsive transcriptional programs and chaperone-dependent regulation of client proteins.

HSPA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSPA2 expression without altering the underlying DNA sequence.

HSPA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSPA2 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 HSPA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSPA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSPA2 locus and enabling the study of HSPA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSPA2 pathway restoration in tumor cells with silenced or reduced HSPA2 expression.

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