αA-crystallin CRISPR/Cas9 KO Plasmid (m): sc-419820

Overview

Cryaa encodes αA-crystallin, a highly abundant small heat shock protein in the ocular lens that functions as an ATP-independent molecular chaperone to maintain crystallin solubility and prevent protein aggregation under stress. Beyond its structural role in establishing lens transparency and refractive properties, αA-crystallin contributes to proteostasis by interacting with misfolded client proteins and modulating stress-response networks linked to redox balance and apoptosis. Altered CRYAA expression or function is associated with lens fiber cell dysfunction, disrupted protein homeostasis, and cataract-related phenotypes, making it a key node for studying lens development and age-related protein aggregation processes. In mouse systems, Cryaa perturbation supports mechanistic work on chaperone activity, cytoskeletal organization, and stress resilience in lens epithelial and fiber cells.

αA-crystallin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Cryaa gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Cryaa 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 Cryaa 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 αA-crystallin protein expression.

This CRISPR knockout system enables efficient generation of Cryaa-deficient cell models for investigation of αA-crystallin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting Cryaa exon(s) critical for αA-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 Cryaa genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by αA-crystallin CRISPR/Cas9 KO Plasmid (m) and αA-crystallin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Cryaa locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by αA-crystallin HDR Plasmid (m) and αA-crystallin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Cryaa homology arms to support homology-directed repair at defined Cryaa 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.