NFKBIA/IkB alpha CRISPR/Cas9 KO Plasmid (m): sc-421878

Overview

Nfkbia encodes NF-κB inhibitor alpha (IκBα), a key negative regulator of canonical NF-κB signaling that binds RelA/p50 complexes and retains them in the cytoplasm under basal conditions. Upon stimulation by pro-inflammatory cytokines, Toll-like receptor ligands, or antigen receptor signaling, IκBα is phosphorylated by the IKK complex and degraded, enabling NF-κB nuclear translocation and transcription of immune, stress-response, and survival genes. This feedback circuitry shapes innate and adaptive immune activation, cytokine production, and cell fate decisions, and is widely used as a mechanistic entry point for studying inflammation-driven tissue remodeling and oncogenic signaling. In mouse systems, perturbation of NFKBIA is commonly leveraged to dissect pathway cross-talk with MAPK, interferon, and apoptotic programs in models of immune dysregulation and tumor microenvironment signaling.

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

This CRISPR knockout system enables efficient generation of Nfkbia-deficient cell models for investigation of NFKBIA/IkB alpha signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting Nfkbia exon(s) critical for NFKBIA/IkB alpha 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 Nfkbia genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

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