NF45 CRISPR Activation Plasmid (h): sc-402042-ACT

ILF2 encodes NF45, a multifunctional RNA- and DNA-binding protein that forms a heterodimer with ILF3 (NF90/NF110) to regulate transcription, pre-mRNA processing, mRNA stability, and translation. NF45 participates in RNA polymerase II–associated gene regulation, stress-responsive ribonucleoprotein complexes, and cellular proliferation programs, and has been linked to control of cytokine and antiviral gene expression. Through these activities, ILF2/NF45 influences cell-cycle progression, genome maintenance, and responses to inflammatory cues. Dysregulation of ILF2 expression or NF45-containing complexes has been reported in contexts of oncogenic growth, immune signaling imbalance, and virus–host interactions, making it a useful node for mechanistic studies of gene regulatory networks.

NF45 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ILF2 expression without altering the underlying DNA sequence.

NF45 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ILF2 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 ILF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NF45 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ILF2 locus and enabling the study of NF45-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NF45 pathway restoration in tumor cells with silenced or reduced ILF2 expression.

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