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.
Cytoplasmic CysRS CRISPR/Cas9 KO Plasmid (h): sc-406444
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cytoplasmic CysRS 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 Cytoplasmic CysRS 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: Cytoplasmic CysRS Antibody (A-3): sc-390230
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cytoplasmic CysRS CRISPR/Cas9 KO Plasmid (h) | sc-406444 | 20 µg | $397.00 |
Overview
CARS encodes cytoplasmic cysteinyl‑tRNA synthetase (CysRS), a class I aminoacyl‑tRNA synthetase that ligates cysteine to its cognate tRNA to ensure translational fidelity in the cytosol. By controlling the supply of charged tRNA\(^\text{Cys}\), cytoplasmic CysRS supports global protein synthesis and couples amino acid availability to proteostasis pathways, including stress responses that modulate translation. Perturbation of aminoacyl‑tRNA synthetase activity can alter ribosome dynamics and trigger integrated stress signaling, with downstream effects on cell growth, metabolism, and redox-sensitive processes that depend on cysteine utilization. Dysregulation of tRNA charging and translation quality control is relevant to research on neurodevelopmental and neuromuscular phenotypes, proliferative disorders, and cellular stress adaptation.
Cytoplasmic CysRS CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CARS gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CARS 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 CARS 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 Cytoplasmic CysRS protein expression.
This CRISPR knockout system enables efficient generation of CARS-deficient cell models for investigation of Cytoplasmic CysRS signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CARS exon(s) critical for Cytoplasmic CysRS 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 CARS genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Cytoplasmic CysRS CRISPR/Cas9 KO Plasmid (h) and Cytoplasmic CysRS CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CARS locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Cytoplasmic CysRS HDR Plasmid (h) and Cytoplasmic CysRS HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CARS homology arms to support homology-directed repair at defined CARS 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.