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.
Fe65 CRISPR/Cas9 KO Plasmid (h): sc-403226
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Fe65 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 Fe65 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: Fe65 Antibody (F-6): sc-398389
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Fe65 CRISPR/Cas9 KO Plasmid (h) | sc-403226 | 20 µg | $397.00 |
Overview
APBB1 encodes the adaptor protein Fe65, a neuronal-enriched interactor of amyloid precursor protein (APP) that couples membrane trafficking to nuclear signaling. Fe65 participates in APP processing and endocytic pathways through PTB-domain binding to APP and related receptors, and it assembles multiprotein complexes with factors such as AICD and transcriptional regulators to influence gene expression. Through these interactions, Fe65 links synaptic function, cytoskeletal dynamics, and regulated proteolysis, processes frequently interrogated in neurodegeneration research. Altered APP–Fe65 signaling and downstream transcriptional programs have been studied in the context of Alzheimer’s disease–related mechanisms and broader neuronal stress responses.
Fe65 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the APBB1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the APBB1 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 APBB1 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 Fe65 protein expression.
This CRISPR knockout system enables efficient generation of APBB1-deficient cell models for investigation of Fe65 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting APBB1 exon(s) critical for Fe65 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 APBB1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Fe65 CRISPR/Cas9 KO Plasmid (h) and Fe65 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the APBB1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Fe65 HDR Plasmid (h) and Fe65 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by APBB1 homology arms to support homology-directed repair at defined APBB1 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.