BTG1 CRISPR Activation Plasmid (h): sc-405339-ACT

BTG1 (B-cell translocation gene 1) encodes an antiproliferative regulator that modulates cell-cycle progression and cellular differentiation, with prominent roles in controlling growth-related transcriptional programs. BTG1 interacts with transcriptional and post-transcriptional machinery, including CCR4–NOT deadenylase complexes, linking it to mRNA turnover and fine-tuning of gene expression. In immune and hematopoietic contexts, BTG1 contributes to homeostatic control of lymphoid cell expansion and stress-responsive signaling. Altered BTG1 expression or disruption of its regulatory network is frequently studied in relation to oncogenic processes, particularly in B-cell malignancy biology and related transcriptional dysregulation.

BTG1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BTG1 expression without altering the underlying DNA sequence.

BTG1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BTG1 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 BTG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BTG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BTG1 locus and enabling the study of BTG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BTG1 pathway restoration in tumor cells with silenced or reduced BTG1 expression.

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