γF-crystallin CRISPR/Cas9 KO Plasmid (m): sc-419833
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- γF-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 γF-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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
γF-crystallin CRISPR/Cas9 KO Plasmid (m) | sc-419833 | 20 µg | $397.00 |
Overview
Crygf encodes γF-crystallin, a highly abundant structural crystallin in the mouse ocular lens that supports refractive index, transparency, and long-term fiber cell stability. γ-crystallins are characterized by tightly packed β-sheet domains that contribute to high protein density while minimizing light scattering, and their maintenance depends on lens proteostasis networks including chaperone interactions, redox balance, and limited protein turnover. Perturbation of crystallin composition or solubility can disrupt lens architecture, promote protein aggregation, and increase susceptibility to cataract-like opacity phenotypes in experimental models. Crygf therefore serves as a useful node for studying lens development, fiber cell differentiation, and mechanisms of protein stability in a largely post-mitotic tissue context.
γF-crystallin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Crygf gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Crygf 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 Crygf 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 γF-crystallin protein expression.
This CRISPR knockout system enables efficient generation of Crygf-deficient cell models for investigation of γF-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Crygf exon(s) critical for γF-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 Crygf genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by γF-crystallin CRISPR/Cas9 KO Plasmid (m) and γF-crystallin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Crygf locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by γF-crystallin HDR Plasmid (m) and γF-crystallin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Crygf homology arms to support homology-directed repair at defined Crygf 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.