OPN1SW CRISPR Activation Plasmid (h): sc-400755-ACT

OPN1SW encodes short-wavelength–sensitive opsin (S-opsin), a G protein–coupled receptor localized to cone photoreceptor outer segments that mediates blue-light detection. Upon photon absorption, OPN1SW activates transducin-dependent phototransduction, driving cGMP phosphodiesterase signaling, closure of cGMP-gated ion channels, and downstream changes in membrane potential that shape visual signaling. Proper OPN1SW expression and chromophore binding are required for cone function, spectral tuning, and retinal circuit integration. Altered opsin activity or cone photoreceptor dysfunction is relevant to inherited color vision deficiencies and broader retinal degenerative processes, supporting mechanistic studies of cone biology.

OPN1SW CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OPN1SW expression without altering the underlying DNA sequence.

OPN1SW CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OPN1SW 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 OPN1SW transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OPN1SW expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OPN1SW locus and enabling the study of OPN1SW-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OPN1SW pathway restoration in tumor cells with silenced or reduced OPN1SW expression.

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