CNBP CRISPR Activation Plasmid (h): sc-404090-ACT

Human CNBP (cellular nucleic acid-binding protein), also known as zinc finger protein 9, is a conserved single-stranded nucleic acid–binding factor that modulates transcription and translation of GC-rich targets. CNBP participates in RNA metabolism and ribonucleoprotein complex dynamics, influencing processes such as stress-responsive gene regulation, cell growth, and differentiation. Through interactions with regulatory nucleic acids and promoter elements, CNBP can shape programs linked to proteostasis and developmental signaling. Dysregulated CNBP activity and repeat-associated mechanisms have been implicated in neuromuscular and neurodevelopmental disease biology, supporting its use in studies of gene expression control and pathomechanisms.

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

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

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