FAST-1 CRISPR/Cas9 KO Plasmid (h): sc-406686
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FAST-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 FAST-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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FAST-1 CRISPR/Cas9 KO Plasmid (h) | sc-406686 | 20 µg | $397.00 |
Overview
FOXH1 encodes the forkhead box transcription factor FAST-1, a nuclear mediator of TGF-β superfamily signaling that cooperates with SMAD2/3-SMAD4 complexes to regulate NODAL/Activin-responsive transcription. FAST-1 binds forkhead motifs in target promoters and integrates developmental cues controlling mesendoderm specification, left–right patterning, and epithelial–mesenchymal transitions through context-dependent transcriptional programs. Dysregulated FOXH1 activity has been linked to congenital laterality defects and perturbed early embryonic development, and altered TGF-β/SMAD transcriptional output is frequently associated with oncogenic processes such as invasion and metastasis. In cell models, FOXH1 provides a tractable node for dissecting SMAD cofactor usage, chromatin recruitment, and pathway cross-talk affecting differentiation and tumor cell plasticity.
FAST-1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the FOXH1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the FOXH1 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 FOXH1 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 FAST-1 protein expression.
This CRISPR knockout system enables efficient generation of FOXH1-deficient cell models for investigation of FAST-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting FOXH1 exon(s) critical for FAST-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 FOXH1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by FAST-1 CRISPR/Cas9 KO Plasmid (h) and FAST-1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the FOXH1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by FAST-1 HDR Plasmid (h) and FAST-1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by FOXH1 homology arms to support homology-directed repair at defined FOXH1 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.