α-actinin-4 CRISPR/Cas9 KO Plasmid (h2): sc-400548-KO-2

Overview

ACTN4 encodes α-actinin-4, a calcium-insensitive actin filament crosslinking protein that localizes to focal adhesions, stress fibers, and cell–cell junctions to regulate cytoskeletal architecture and mechanical signaling. By organizing F-actin networks and linking them to integrin-associated complexes, α-actinin-4 contributes to cell adhesion, spreading, migration, and changes in cell stiffness that influence pathways such as Rho GTPase signaling and mechanotransduction. In addition to its cytoplasmic roles, α-actinin-4 has been implicated in nuclear functions including modulation of transcriptional programs through protein–protein interactions. Dysregulated ACTN4 expression or mutation has been associated with kidney podocyte dysfunction (including focal segmental glomerulosclerosis) and with invasive phenotypes in multiple tumor types, supporting its relevance for studies of cytoskeletal control and disease-linked cell behavior.

α-actinin-4 CRISPR/Cas9 KO Plasmid (h2) is a pool of plasmids designed for targeted disruption of the ACTN4 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ACTN4 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 ACTN4 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 α-actinin-4 protein expression.

This CRISPR knockout system enables efficient generation of ACTN4-deficient cell models for investigation of α-actinin-4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting ACTN4 exon(s) critical for α-actinin-4 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 ACTN4 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by α-actinin-4 CRISPR/Cas9 KO Plasmid (h) and α-actinin-4 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ACTN4 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by α-actinin-4 HDR Plasmid (h) and α-actinin-4 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ACTN4 homology arms to support homology-directed repair at defined ACTN4 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.