NF90 CRISPR Activation Plasmid (h): sc-402626-ACT

ILF3 encodes NF90, a multifunctional double-stranded RNA-binding protein that regulates gene expression at the transcriptional and post-transcriptional levels through control of mRNA stability, splicing, transport, and translation. NF90 participates in RNA processing and stress-responsive programs, including antiviral signaling and inflammatory gene regulation, and it can modulate cell-cycle progression via interactions with transcriptional and RNA maturation machinery. Through these roles, ILF3/NF90 influences pathways such as interferon-stimulated gene expression, RNA surveillance, and ribonucleoprotein complex assembly. Dysregulated NF90 activity and altered ILF3 expression have been reported in contexts relevant to cancer biology, immune-mediated inflammation, and viral infection, supporting its use as a mechanistic node in gene regulation studies.

NF90 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ILF3 expression without altering the underlying DNA sequence.

NF90 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ILF3 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 ILF3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NF90 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ILF3 locus and enabling the study of NF90-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NF90 pathway restoration in tumor cells with silenced or reduced ILF3 expression.

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