Che-1 CRISPR Activation Plasmid (h): sc-404982-ACT

Human AATF encodes Che-1, a nuclear transcriptional cofactor implicated in coordinating cell-cycle progression, DNA damage signaling, and apoptotic thresholds. Che-1 interacts with transcriptional and chromatin-regulatory machinery to modulate expression programs linked to p53-dependent stress responses and RNA polymerase II–mediated transcription. Through these functions, AATF contributes to cellular decisions between proliferation and checkpoint arrest, and its dysregulation has been associated with altered survival signaling and genomic instability phenotypes in cancer-relevant contexts. Che-1 activity is also studied in relation to proteostasis and RNA metabolism, supporting its use in mechanistic investigations of stress adaptation pathways.

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

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

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