BTG2 CRISPR Activation Plasmid (m): sc-419374-ACT

Btg2 (BTG2) is an antiproliferative, immediate-early regulator that modulates cell-cycle progression, differentiation, and stress-responsive transcription in mouse cells. BTG2 interfaces with transcriptional control and mRNA turnover pathways, including interactions with CCR4–NOT deadenylase complexes, linking mitogenic signals to changes in gene expression and cellular growth restraint. It is frequently studied in the context of DNA damage signaling and p53-associated responses, where altered BTG2 activity can influence checkpoint control, apoptosis susceptibility, and cellular senescence programs. Dysregulated BTG2 expression has been associated with oncogenic transformation and tumor progression in multiple model systems, making it relevant for mechanistic studies of proliferation control and cancer-associated transcriptional networks.

BTG2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Btg2 expression without altering the underlying DNA sequence.

BTG2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Btg2 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 Btg2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BTG2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Btg2 locus and enabling the study of BTG2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BTG2 pathway restoration in tumor cells with silenced or reduced Btg2 expression.

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