nucleobindin 2 CRISPR Activation Plasmid (m): sc-424686-ACT

Mouse Nucb2 encodes nucleobindin 2, a Ca2+-binding, EF-hand–containing protein that localizes to secretory pathway compartments and participates in regulated secretion and cellular Ca2+ homeostasis. Nucleobindin 2 has been linked to neuroendocrine signaling through processing into nesfatin-1–related peptides, with reported roles in appetite/energy balance and stress-associated responses. In peripheral tissues, altered Nucb2 expression has been studied in the context of metabolic regulation, inflammation-associated signaling, and cellular adaptation to nutrient status. These properties make Nucb2 a useful target for dissecting Ca2+-dependent trafficking, secretory granule biology, and neuroendocrine-metabolic crosstalk in mouse models.

nucleobindin 2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nucb2 expression without altering the underlying DNA sequence.

nucleobindin 2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nucb2 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 Nucb2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous nucleobindin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nucb2 locus and enabling the study of nucleobindin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of nucleobindin 2 pathway restoration in tumor cells with silenced or reduced Nucb2 expression.

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