γC-crystallin CRISPR/Cas9 KO Plasmid (h): sc-402940
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- γC-crystallin 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 γC-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 | |
γC-crystallin CRISPR/Cas9 KO Plasmid (h) | sc-402940 | 20 µg | $397.00 |
Overview
CRYGC encodes human γC-crystallin, a highly abundant structural protein in the ocular lens that contributes to lens transparency and refractive properties by maintaining densely packed, soluble protein assemblies. As a member of the γ-crystallin family, it supports long-term protein stability in terminally differentiated lens fiber cells that lack protein turnover machinery. Disruption of crystallin homeostasis promotes protein misfolding and aggregation, processes linked to loss of lens clarity. Genetic variation or altered expression of CRYGC has been associated with inherited cataract phenotypes, making it a useful target for studying proteostasis and lens development.
γC-crystallin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CRYGC gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CRYGC 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 CRYGC 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 γC-crystallin protein expression.
This CRISPR knockout system enables efficient generation of CRYGC-deficient cell models for investigation of γC-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CRYGC exon(s) critical for γC-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 CRYGC genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by γC-crystallin CRISPR/Cas9 KO Plasmid (h) and γC-crystallin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CRYGC locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by γC-crystallin HDR Plasmid (h) and γC-crystallin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CRYGC homology arms to support homology-directed repair at defined CRYGC 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.