BMAL1 CRISPR Activation Plasmid (m): sc-419206-ACT

Arntl encodes BMAL1, a core basic helix–loop–helix PAS transcription factor that heterodimerizes with CLOCK to drive circadian gene expression programs in mouse tissues. BMAL1 regulates rhythmic transcription of clock-controlled genes involved in metabolism, cell cycle control, DNA damage responses, and immune signaling, integrating environmental cues with cellular physiology. Through modulation of transcriptional feedback loops involving PER and CRY proteins, BMAL1 influences mitochondrial function, oxidative stress handling, and endocrine homeostasis. Dysregulated BMAL1 activity has been associated with disrupted circadian rhythms and related phenotypes in models of metabolic dysfunction, inflammation, and tumor biology, supporting its utility in mechanistic research.

BMAL1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Arntl expression without altering the underlying DNA sequence.

BMAL1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Arntl 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 Arntl transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BMAL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Arntl locus and enabling the study of BMAL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BMAL1 pathway restoration in tumor cells with silenced or reduced Arntl expression.

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