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.
IDE CRISPR/Cas9 KO Plasmid (m): sc-421021
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- IDE 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 IDE 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: IDE Antibody (F-9): sc-393887
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IDE CRISPR/Cas9 KO Plasmid (m) | sc-421021 | 20 µg | $397.00 |
Overview
Insulin degrading enzyme (IDE) is a zinc metalloprotease that mediates intracellular and extracellular proteolysis of insulin and other small peptides, contributing to peptide turnover and regulation of signaling duration. In mouse tissues, IDE activity interfaces with insulin/IGF signaling, endosomal trafficking, and proteostasis pathways that shape metabolic homeostasis and stress responses. Altered IDE expression or function has been linked to dysregulated glucose handling and peptide clearance, and it is frequently investigated in models relevant to metabolic phenotypes and neurodegeneration-associated peptide accumulation. As a result, Ide is a useful genetic node for dissecting how peptide degradation influences cellular signaling networks and aggregate-prone protein biology.
IDE CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ide gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ide 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 Ide 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 IDE protein expression.
This CRISPR knockout system enables efficient generation of Ide-deficient cell models for investigation of IDE signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ide exon(s) critical for IDE 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 Ide genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by IDE CRISPR/Cas9 KO Plasmid (m) and IDE CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ide locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by IDE HDR Plasmid (m) and IDE HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ide homology arms to support homology-directed repair at defined Ide 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.