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.
API5 CRISPR/Cas9 KO Plasmid (h): sc-406608
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- API5 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 API5 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: API5 Antibody (E-12): sc-374528
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
API5 CRISPR/Cas9 KO Plasmid (h) | sc-406608 | 20 µg | $397.00 |
Overview
API5 (apoptosis inhibitor 5) encodes a nuclear protein that contributes to cell survival by modulating apoptotic signaling and supporting cellular stress tolerance. API5 has been linked to regulation of E2F-dependent transcriptional programs, cell-cycle progression, and maintenance of proliferative capacity, with reported interactions that influence chromatin-associated processes. Altered API5 expression is frequently studied in the context of oncogenic phenotypes, including enhanced resistance to apoptosis and changes in growth control. As a result, API5 is a useful target for dissecting mechanisms of survival signaling, transcriptional regulation, and tumor-associated cell-state transitions.
API5 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the API5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the API5 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 API5 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 API5 protein expression.
This CRISPR knockout system enables efficient generation of API5-deficient cell models for investigation of API5 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting API5 exon(s) critical for API5 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 API5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by API5 CRISPR/Cas9 KO Plasmid (h) and API5 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the API5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by API5 HDR Plasmid (h) and API5 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by API5 homology arms to support homology-directed repair at defined API5 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.