Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
AIRE-1 CRISPR/Cas9 KO Plasmid (h): sc-401225
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AIRE-1 CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 AIRE-1 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: AIRE-1 Antibody (C-2): sc-373703
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AIRE-1 CRISPR/Cas9 KO Plasmid (h) | sc-401225 | 20 µg | $397.00 |
Overview
Autoimmune regulator (AIRE; AIRE-1) is a nuclear transcriptional regulator that enforces central immune tolerance by promoting ectopic expression of tissue-restricted antigens in medullary thymic epithelial cells, enabling negative selection and shaping the T cell repertoire. AIRE coordinates chromatin- and transcription-associated processes through interactions with histone marks and transcriptional machinery, supporting gene expression programs linked to thymic selection and immune homeostasis. Disruption of AIRE function is associated with breakdown of tolerance and autoimmunity, and altered AIRE-dependent transcriptional programs have been studied in the context of inflammatory phenotypes and immune dysregulation. As a result, AIRE is widely used as a model locus for investigating transcriptional control, antigen presentation biology, and mechanisms governing self-tolerance.
AIRE-1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the AIRE gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the AIRE 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 AIRE 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 AIRE-1 protein expression.
This CRISPR knockout system enables efficient generation of AIRE-deficient cell models for investigation of AIRE-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting AIRE exon(s) critical for AIRE-1 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 AIRE genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by AIRE-1 CRISPR/Cas9 KO Plasmid (h) and AIRE-1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the AIRE locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by AIRE-1 HDR Plasmid (h) and AIRE-1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by AIRE homology arms to support homology-directed repair at defined AIRE 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.