Ataxin-1 CRISPR Activation Plasmid (h): sc-403309-ACT

ATXN1 encodes ataxin-1, a predominantly nuclear protein that participates in transcriptional regulation and RNA-associated processes through interactions with chromatin and RNA-binding partners. Ataxin-1 contributes to neuronal and developmental gene expression programs and is linked to protein homeostasis pathways, including ubiquitin-dependent turnover and subnuclear compartmentalization. Altered ATXN1 dosage or function can perturb transcriptional networks and cellular stress responses, and the gene is widely studied in the context of neurodegeneration, particularly polyglutamine-associated spinocerebellar ataxia. In human cell models, ATXN1 serves as a tractable node for dissecting gene regulatory circuitry, proteostasis mechanisms, and cell-type-specific vulnerability.

Ataxin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATXN1 expression without altering the underlying DNA sequence.

Ataxin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATXN1 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 ATXN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ataxin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATXN1 locus and enabling the study of Ataxin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ataxin-1 pathway restoration in tumor cells with silenced or reduced ATXN1 expression.

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