Bmi-1 CRISPR Activation Plasmid (h): sc-417606-ACT

BMI1 encodes the Polycomb group protein Bmi-1, a core component of Polycomb repressive complex 1 (PRC1) that maintains epigenetic gene silencing through chromatin compaction and H2A ubiquitination. Bmi-1 regulates transcriptional programs controlling self-renewal, lineage commitment, and cell-cycle progression, including repression of the CDKN2A/INK4A–ARF locus and modulation of senescence checkpoints. Through these functions, BMI1 influences DNA damage responses, oxidative stress tolerance, and stem-like phenotypes in diverse cell contexts. Dysregulated BMI1 activity has been widely associated with oncogenic transcriptional states and altered differentiation in multiple disease-relevant models, making it a frequent target in epigenetics and cancer biology research.

Bmi-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BMI1 expression without altering the underlying DNA sequence.

Bmi-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BMI1 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 BMI1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Bmi-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BMI1 locus and enabling the study of Bmi-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Bmi-1 pathway restoration in tumor cells with silenced or reduced BMI1 expression.

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