Nrl CRISPR Activation Plasmid (h): sc-402610-ACT

NRL (neural retina leucine zipper) encodes the transcription factor Nrl, a basic leucine zipper regulator that plays a central role in photoreceptor development and the establishment of rod cell identity. By binding cis-regulatory elements and coordinating transcriptional networks with other retinal factors, Nrl controls programs involved in phototransduction, outer segment formation, and metabolic homeostasis in differentiated neurons. Dysregulation of NRL-associated gene expression is linked to inherited retinal disorders and impaired photoreceptor function, making NRL a useful node for studying gene regulatory circuitry in retinal biology. Human Nrl activity is therefore relevant to investigations of neuronal differentiation, cell fate specification, and transcriptional control in the retina.

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

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

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