REP-2 CRISPR/Cas9 KO Plasmid (h): sc-417782
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- REP-2 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 REP-2 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: REP-2 Antibody (E-4): sc-398605
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
REP-2 CRISPR/Cas9 KO Plasmid (h) | sc-417782 | 20 µg | $397.00 |
Overview
CHML encodes Rab escort protein 2 (REP-2), a cytosolic chaperone that binds newly synthesized Rab GTPases and presents them to Rab geranylgeranyltransferase for prenylation, a lipid modification required for membrane association. Through this Rab maturation step, REP-2 supports vesicular trafficking pathways including endosome and Golgi transport, exocytosis, and autophagy-related membrane dynamics. Disruption of Rab prenylation or Rab cycling can perturb organelle identity and intracellular cargo sorting, processes frequently implicated in neurodegeneration, retinal disorders, and broader trafficking-related cellular stress phenotypes. CHML is also studied in contexts where Rab-dependent signaling and membrane remodeling intersect with immune cell function and cancer cell biology.
REP-2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CHML gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CHML 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 CHML 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 REP-2 protein expression.
This CRISPR knockout system enables efficient generation of CHML-deficient cell models for investigation of REP-2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CHML exon(s) critical for REP-2 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 CHML genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by REP-2 CRISPR/Cas9 KO Plasmid (h) and REP-2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CHML locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by REP-2 HDR Plasmid (h) and REP-2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CHML homology arms to support homology-directed repair at defined CHML 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.