βB3-crystallin CRISPR/Cas9 KO Plasmid (m): sc-419827
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- βB3-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 βB3-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: βB3-crystallin Antibody (G-3): sc-374374
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
βB3-crystallin CRISPR/Cas9 KO Plasmid (m) | sc-419827 | 20 µg | $397.00 |
Overview
Crybb3 encodes βB3-crystallin, a structural β/γ-crystallin family protein that contributes to lens fiber cell architecture, long-range order, and maintenance of transparency. βB3-crystallin participates in crystallin complex assembly and supports protein stability and refractive properties during lens development and maturation. Disruption of crystallin homeostasis can promote protein aggregation, altered redox balance, and lens light-scattering, linking Crybb3 to pathways governing proteostasis and cellular stress responses in the ocular lens. In mouse models, perturbation of crystallin composition is commonly studied in the context of lens degeneration and cataract-like phenotypes.
βB3-crystallin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Crybb3 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Crybb3 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 Crybb3 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 βB3-crystallin protein expression.
This CRISPR knockout system enables efficient generation of Crybb3-deficient cell models for investigation of βB3-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Crybb3 exon(s) critical for βB3-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 Crybb3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by βB3-crystallin CRISPR/Cas9 KO Plasmid (m) and βB3-crystallin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Crybb3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by βB3-crystallin HDR Plasmid (m) and βB3-crystallin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Crybb3 homology arms to support homology-directed repair at defined Crybb3 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.