Brn-3a/BRN3A/POU4F1 CRISPR Activation Plasmid (h): sc-400322-ACT

POU4F1 (Brn-3a/BRN3A) encodes a POU-domain transcription factor that regulates neuronal lineage specification, survival, and terminal differentiation by controlling gene programs involved in axon guidance, neurite outgrowth, and stress-responsive transcription. It is highly associated with sensory neuron development and is frequently used as a marker and mechanistic driver in studies of neural crest-derived and peripheral neuronal cell states. Brn-3a interfaces with transcriptional networks that modulate apoptosis and cell-cycle control through downstream regulation of pro-survival and differentiation-linked targets. Dysregulated POU4F1 activity has been reported in contexts of neurodevelopmental dysfunction and oncogenic transcriptional reprogramming, supporting its utility for investigating disease-relevant gene regulation.

Brn-3a/BRN3A/POU4F1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous POU4F1 expression without altering the underlying DNA sequence.

Brn-3a/BRN3A/POU4F1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the POU4F1 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 POU4F1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Brn-3a/BRN3A/POU4F1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native POU4F1 locus and enabling the study of Brn-3a/BRN3A/POU4F1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Brn-3a/BRN3A/POU4F1 pathway restoration in tumor cells with silenced or reduced POU4F1 expression.

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