FAST CRISPR/Cas9 KO Plasmid (h): sc-406169
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FAST 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 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: FAST Antibody (B-9): sc-365125
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FAST CRISPR/Cas9 KO Plasmid (h) | sc-406169 | 20 µg | $397.00 |
Overview
FASTK (Fas-activated serine/threonine kinase) encodes a mitochondrial RNA-binding protein implicated in post-transcriptional regulation of mitochondrial gene expression, including processing and stability of specific mtRNAs required for oxidative phosphorylation. Through its roles in mitochondrial homeostasis, FASTK influences cellular bioenergetics, stress adaptation, and apoptotic signaling linked to Fas-mediated pathways. Dysregulation of mitochondrial RNA metabolism and OXPHOS capacity associated with FASTK has been examined in contexts of metabolic stress, neurodegeneration, and cancer cell survival phenotypes where mitochondrial function is a key determinant.
FAST CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the FASTK gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the FASTK 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 FASTK 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 protein expression.
This CRISPR knockout system enables efficient generation of FASTK-deficient cell models for investigation of FAST signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting FASTK exon(s) critical for FAST 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 FASTK genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by FAST CRISPR/Cas9 KO Plasmid (h) and FAST CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the FASTK locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by FAST HDR Plasmid (h) and FAST HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by FASTK homology arms to support homology-directed repair at defined FASTK 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.