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.
AFP/Alpha fetoprotein CRISPR/Cas9 KO Plasmid (h): sc-400355
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AFP/Alpha fetoprotein 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 AFP/Alpha fetoprotein 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: AFP/Alpha fetoprotein Antibody (C3): sc-8399
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AFP/Alpha fetoprotein CRISPR/Cas9 KO Plasmid (h) | sc-400355 | 20 µg | $397.00 |
Overview
AFP (alpha-fetoprotein) encodes a secreted oncofetal glycoprotein that is highly expressed during fetal development and largely silenced after birth, where it contributes to regulation of growth and differentiation programs. In adult tissues, re-expression of AFP is frequently associated with altered hepatic lineage states and remodeling of extracellular and immune signaling in the tumor microenvironment. AFP expression is used as a molecular feature in studies of hepatocellular carcinoma and germ cell tumor biology, and it is commonly evaluated alongside pathways linked to hepatocyte differentiation, proliferation, and stress responses. As a circulating protein, AFP also serves as a tractable readout for transcriptional regulation and secretory pathway dynamics in human cell models.
AFP/Alpha fetoprotein CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the AFP gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the AFP 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 AFP 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 AFP/Alpha fetoprotein protein expression.
This CRISPR knockout system enables efficient generation of AFP-deficient cell models for investigation of AFP/Alpha fetoprotein signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting AFP exon(s) critical for AFP/Alpha fetoprotein 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 AFP genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by AFP/Alpha fetoprotein CRISPR/Cas9 KO Plasmid (h) and AFP/Alpha fetoprotein CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the AFP locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by AFP/Alpha fetoprotein HDR Plasmid (h) and AFP/Alpha fetoprotein HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by AFP homology arms to support homology-directed repair at defined AFP 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.