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.
ValRS CRISPR/Cas9 KO Plasmid (m): sc-423653
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ValRS CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 ValRS 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: ValRS Antibody (D-7): sc-166674
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ValRS CRISPR/Cas9 KO Plasmid (m) | sc-423653 | 20 µg | $397.00 |
Overview
Vars encodes valyl‑tRNA synthetase (ValRS), a cytosolic aminoacyl‑tRNA synthetase that ligates valine to its cognate tRNA, ensuring translational fidelity and proteome homeostasis. By maintaining accurate decoding during protein synthesis, ValRS supports core processes including ribosome function, integrated stress responses to amino acid imbalance, and broader proteostasis networks. Perturbation of aminoacyl‑tRNA charging can disrupt cell growth and stress adaptation, linking this pathway to mechanisms relevant to neurodevelopment, neuromuscular biology, and mitochondrial–cytosolic coordination via altered proteome demand.
ValRS CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Vars gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Vars 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 Vars 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 ValRS protein expression.
This CRISPR knockout system enables efficient generation of Vars-deficient cell models for investigation of ValRS signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Vars exon(s) critical for ValRS 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 Vars genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ValRS CRISPR/Cas9 KO Plasmid (m) and ValRS CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Vars locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ValRS HDR Plasmid (m) and ValRS HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Vars homology arms to support homology-directed repair at defined Vars 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.