AIP4 CRISPR/Cas9 KO Plasmid (m): sc-421158
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AIP4 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 AIP4 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AIP4 CRISPR/Cas9 KO Plasmid (m) | sc-421158 | 20 µg | $397.00 |
Overview
Itch (AIP4) encodes a HECT-type E3 ubiquitin ligase that catalyzes ubiquitination of target proteins to control their stability and trafficking, shaping proteostasis and signal duration. In mouse cells, AIP4 participates in ubiquitin-dependent regulation of immune signaling pathways, including modulation of NF-κB and related receptor-proximal complexes, thereby influencing cytokine outputs and inflammatory homeostasis. Itch also contributes to T cell differentiation and peripheral tolerance through turnover of transcriptional regulators and signaling adaptors. Dysregulated Itch activity has been linked to immune imbalance, inflammatory phenotypes, and altered control of cell growth and survival pathways relevant to cancer biology and neuroimmune interactions.
AIP4 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Itch gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Itch 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 Itch 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 AIP4 protein expression.
This CRISPR knockout system enables efficient generation of Itch-deficient cell models for investigation of AIP4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Itch exon(s) critical for AIP4 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 Itch genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by AIP4 CRISPR/Cas9 KO Plasmid (m) and AIP4 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Itch locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by AIP4 HDR Plasmid (m) and AIP4 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Itch homology arms to support homology-directed repair at defined Itch 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.