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.
γB-crystallin CRISPR/Cas9 KO Plasmid (h): sc-402939
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- γB-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 γB-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: γB-crystallin Antibody (D-5): sc-377056
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
γB-crystallin CRISPR/Cas9 KO Plasmid (h) | sc-402939 | 20 µg | $397.00 |
Overview
CRYGB encodes γB-crystallin, a highly abundant, soluble structural protein of the vertebrate eye lens that contributes to refractive index and long-term lens transparency. γB-crystallin participates in crystallin network assembly and protein homeostasis processes that maintain lens fiber cell integrity, where tight packing and stability are essential to minimize light scattering. Disruption of crystallin stability or increased susceptibility to aggregation is linked to lens opacification phenotypes, making CRYGB a relevant locus for studying mechanisms of protein misfolding and proteostasis in differentiated ocular tissues. Because lens fiber cells are largely post-mitotic, CRYGB also provides a model for investigating how long-lived proteins respond to oxidative and environmental stress over time.
γB-crystallin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CRYGB gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CRYGB 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 CRYGB 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 γB-crystallin protein expression.
This CRISPR knockout system enables efficient generation of CRYGB-deficient cell models for investigation of γB-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CRYGB exon(s) critical for γB-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 CRYGB genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by γB-crystallin CRISPR/Cas9 KO Plasmid (h) and γB-crystallin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CRYGB locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by γB-crystallin HDR Plasmid (h) and γB-crystallin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CRYGB homology arms to support homology-directed repair at defined CRYGB 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.