HAX-1 CRISPR Activation Plasmid (h): sc-402087-ACT

HAX1 encodes HAX-1, a predominantly mitochondrial and endoplasmic reticulum–associated protein that regulates apoptosis, calcium homeostasis, and cytoskeletal dynamics. It participates in stress-response and survival signaling by modulating mitochondrial membrane integrity and interacting with BCL-2 family–related processes, influencing caspase activation and reactive oxygen species balance. HAX-1 also contributes to actin remodeling and intracellular trafficking, linking it to cell migration and adhesion programs. Dysregulated HAX1 expression or function has been associated with altered hematopoietic cell survival, immune cell homeostasis, and tumor biology, making it a useful target for studying survival pathways and cell fate decisions.

HAX-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HAX1 expression without altering the underlying DNA sequence.

HAX-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HAX1 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 HAX1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HAX-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HAX1 locus and enabling the study of HAX-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HAX-1 pathway restoration in tumor cells with silenced or reduced HAX1 expression.

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