ROS-GC1 CRISPR/Cas9 KO Plasmid (h): sc-404483
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ROS-GC1 CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the ROS-GC1 genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ROS-GC1 Antibody (B-7): sc-376217
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ROS-GC1 CRISPR/Cas9 KO Plasmid (h) | sc-404483 | 20 µg | $397.00 |
Overview
GUCY2D encodes ROS-GC1, a membrane-bound receptor guanylate cyclase that synthesizes cGMP in response to intracellular Ca2+ signals conveyed by guanylate cyclase–activating proteins. In retinal photoreceptors, ROS-GC1 supports recovery and adaptation by reconstituting cGMP levels that control cyclic nucleotide–gated channel activity within the phototransduction cascade. This enzyme integrates calcium homeostasis with second-messenger signaling to tune membrane excitability and synaptic output. Dysregulation or pathogenic variants in GUCY2D are linked to inherited retinal disorders, making it a key target for mechanistic studies of cGMP signaling and sensory neuron function.
ROS-GC1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the GUCY2D gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the GUCY2D together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the GUCY2D open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish ROS-GC1 protein expression.
This CRISPR knockout system enables efficient generation of GUCY2D-deficient cell models for investigation of ROS-GC1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting GUCY2D exon(s) critical for ROS-GC1 function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple GUCY2D genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ROS-GC1 CRISPR/Cas9 KO Plasmid (h) and ROS-GC1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the GUCY2D locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ROS-GC1 HDR Plasmid (h) and ROS-GC1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by GUCY2D homology arms to support homology-directed repair at defined GUCY2D target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.