BMAL2 CRISPR Activation Plasmid (m): sc-435462-ACT

Arntl2 encodes the basic helix–loop–helix PAS transcription factor BMAL2, a core regulator of circadian transcriptional programs that heterodimerizes with CLOCK/NPAS2 to drive E-box–dependent gene expression. In mouse cells, BMAL2 contributes to rhythmic control of metabolism, mitochondrial function, DNA damage responses, and immune signaling through clock-controlled networks. Arntl2-mediated timing cues intersect with pathways such as MAPK and NF-κB signaling and shape cell-state transitions under stress. Dysregulated ARNTL2/BMAL2 activity has been associated with altered circadian homeostasis and has been studied in contexts including inflammation, metabolic dysfunction, and tumor-associated phenotypes.

BMAL2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Arntl2 expression without altering the underlying DNA sequence.

BMAL2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Arntl2 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 Arntl2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BMAL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Arntl2 locus and enabling the study of BMAL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BMAL2 pathway restoration in tumor cells with silenced or reduced Arntl2 expression.

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