RFC-1 CRISPR/Cas9 KO Plasmid (m): sc-422982
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RFC-1 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 RFC-1 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: SLC19A1 Antibody (D-6): sc-271276
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RFC-1 CRISPR/Cas9 KO Plasmid (m) | sc-422982 | 20 µg | $397.00 |
Overview
Slc19a1 encodes the reduced folate carrier RFC-1, a major plasma membrane transporter responsible for cellular uptake of reduced folates and related one-carbon metabolites. By controlling intracellular folate availability, RFC-1 supports nucleotide biosynthesis, methylation reactions, and redox homeostasis, linking the transporter to core processes such as DNA replication and epigenetic regulation. Altered Slc19a1 activity can shift folate-dependent metabolic flux and influence proliferative capacity, genome stability, and differentiation programs in mammalian cells. In mouse models, RFC-1 function is frequently examined in the context of folate metabolism, developmental biology, and mechanisms underlying folate-sensitive phenotypes.
RFC-1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Slc19a1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Slc19a1 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 Slc19a1 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 RFC-1 protein expression.
This CRISPR knockout system enables efficient generation of Slc19a1-deficient cell models for investigation of RFC-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Slc19a1 exon(s) critical for RFC-1 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 Slc19a1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RFC-1 CRISPR/Cas9 KO Plasmid (m) and RFC-1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Slc19a1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RFC-1 HDR Plasmid (m) and RFC-1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Slc19a1 homology arms to support homology-directed repair at defined Slc19a1 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.