HSPA1L CRISPR Activation Plasmid (h): sc-400479-ACT

HSPA1L encodes a member of the HSP70 family of ATP-dependent molecular chaperones that supports proteostasis by promoting nascent protein folding, preventing aggregation, and facilitating refolding of stress-denatured substrates. HSPA1L participates in the cellular heat shock response and interfaces with co-chaperone networks to regulate protein quality control, including stabilization of signaling proteins and modulation of stress-adaptive transcriptional programs. Through these functions, HSPA1L can influence pathways linked to endoplasmic reticulum stress, proteasome-mediated turnover, and cell survival decisions during proteotoxic stress. Altered chaperone activity and heat shock signaling are associated with disease-relevant phenotypes in cancer biology, neurodegeneration, and inflammatory stress contexts, making HSPA1L a useful target for mechanistic studies of stress resilience and proteome maintenance.

HSPA1L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSPA1L expression without altering the underlying DNA sequence.

HSPA1L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSPA1L 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 HSPA1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSPA1L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSPA1L locus and enabling the study of HSPA1L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSPA1L pathway restoration in tumor cells with silenced or reduced HSPA1L expression.

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