NF-YA CRISPR Activation Plasmid (h): sc-401085-ACT

NFYA encodes the NF-YA subunit of the heterotrimeric NF-Y transcription factor, which binds CCAAT box motifs and coordinates promoter architecture with chromatin state to control basal and inducible transcription. NF-YA contributes to regulation of cell-cycle progression, DNA replication and repair, metabolic gene programs, and stress-responsive pathways through cooperative interactions with other transcription factors and coactivator complexes. Altered NFYA activity or NF-Y target gene networks has been linked to dysregulated proliferation and differentiation, with relevance to oncogenic transcriptional programs and broader gene-expression changes observed across multiple disease contexts. These properties make NFYA a useful node for interrogating promoter-driven regulation, transcriptional circuitry, and pathway rewiring in human cellular models.

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

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

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