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.
Rad54 CRISPR/Cas9 KO Plasmid (h): sc-401750
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rad54 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 Rad54 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: Rad54 Antibody (F-11): sc-374598
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rad54 CRISPR/Cas9 KO Plasmid (h) | sc-401750 | 20 µg | $397.00 |
Overview
RAD54L encodes the human Rad54 DNA-dependent ATPase, a SWI2/SNF2 family chromatin remodeler that cooperates with RAD51 to promote homologous recombination. Rad54 stabilizes and remodels RAD51 presynaptic filaments, stimulates strand invasion, and supports DNA double-strand break repair during replication stress, contributing to genome integrity and cell-cycle checkpoint competence. The protein functions within the homologous recombination and DNA damage response networks, interacting with key repair factors to regulate recombination intermediate processing. Altered RAD54L activity is relevant to chromosomal instability phenotypes frequently investigated in cancer biology and in models of defective DNA repair.
Rad54 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RAD54L gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RAD54L 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 RAD54L 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 Rad54 protein expression.
This CRISPR knockout system enables efficient generation of RAD54L-deficient cell models for investigation of Rad54 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RAD54L exon(s) critical for Rad54 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 RAD54L genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Rad54 CRISPR/Cas9 KO Plasmid (h) and Rad54 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RAD54L locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Rad54 HDR Plasmid (h) and Rad54 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RAD54L homology arms to support homology-directed repair at defined RAD54L 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.