Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
γS-crystallin CRISPR/Cas9 KO Plasmid (m): sc-419834
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- γS-crystallin CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 γS-crystallin 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: γS-crystallin Antibody (A-3): sc-374265
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
γS-crystallin CRISPR/Cas9 KO Plasmid (m) | sc-419834 | 20 µg | $397.00 |
Overview
Crygs encodes mouse γS-crystallin, a highly abundant structural protein of the ocular lens cytoplasm that contributes to lens transparency and refractive properties by stabilizing densely packed protein assemblies. As a member of the β/γ-crystallin superfamily, γS-crystallin supports long-term protein homeostasis in an anucleate environment, where altered folding, aggregation, or post-translational modification can disrupt lens light transmission. Disruption of crystallin balance intersects with proteostasis pathways, including chaperone-assisted folding and stress responses that influence protein solubility over the lifespan. Genetic and biochemical perturbations of crystallins are widely used to model lens opacity phenotypes and to study mechanisms underlying cataract-associated protein aggregation.
γS-crystallin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Crygs gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Crygs 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 Crygs 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 γS-crystallin protein expression.
This CRISPR knockout system enables efficient generation of Crygs-deficient cell models for investigation of γS-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Crygs exon(s) critical for γS-crystallin 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 Crygs genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by γS-crystallin CRISPR/Cas9 KO Plasmid (m) and γS-crystallin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Crygs locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by γS-crystallin HDR Plasmid (m) and γS-crystallin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Crygs homology arms to support homology-directed repair at defined Crygs 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.