YB-1 CRISPR Activation Plasmid (h): sc-401385-ACT

YBX1 encodes the human Y-box binding protein 1 (YB-1), a multifunctional DNA/RNA-binding factor that regulates transcription, mRNA splicing, translation, and stress granule dynamics. YB-1 participates in cellular stress responses and influences cell-cycle control, apoptosis, and DNA damage signaling through broad control of gene expression programs. It has been linked to pathways governing proliferation and inflammatory signaling, including modulation of oncogenic transcriptional networks and post-transcriptional regulation of growth-related transcripts. Dysregulated YBX1/YB-1 activity is frequently associated with tumor biology, metastasis-related phenotypes, and resistance to cellular stress, making it a key target for mechanistic studies of gene regulation and proteostasis.

YB-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous YBX1 expression without altering the underlying DNA sequence.

YB-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the YBX1 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 YBX1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous YB-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native YBX1 locus and enabling the study of YB-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of YB-1 pathway restoration in tumor cells with silenced or reduced YBX1 expression.

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