MCP-5 CRISPR/Cas9 KO Plasmid (m): sc-422836
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MCP-5 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 MCP-5 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: MCP-5 Antibody (16Q07): sc-74220
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MCP-5 CRISPR/Cas9 KO Plasmid (m) | sc-422836 | 20 µg | $397.00 |
Overview
Mouse Ccl12 encodes monocyte chemoattractant protein-5 (MCP-5), a CC chemokine that promotes chemotaxis of monocytes/macrophages and other leukocyte subsets during inflammatory responses. MCP-5 signals primarily through chemokine receptors such as CCR2 to trigger GPCR-mediated pathways that remodel the cytoskeleton and coordinate directed migration, integrating with broader cytokine and innate immune signaling networks. In vivo and in vitro studies link Ccl12/MCP-5 activity to leukocyte infiltration programs in tissue injury and chronic inflammation, with relevance to fibrosis, metabolic inflammation, and tumor-associated myeloid recruitment. As a secreted chemokine, MCP-5 also serves as a tractable readout for immune cell crosstalk and stromal–immune interactions in mouse models.
MCP-5 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ccl12 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ccl12 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 Ccl12 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 MCP-5 protein expression.
This CRISPR knockout system enables efficient generation of Ccl12-deficient cell models for investigation of MCP-5 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ccl12 exon(s) critical for MCP-5 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 Ccl12 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MCP-5 CRISPR/Cas9 KO Plasmid (m) and MCP-5 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ccl12 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MCP-5 HDR Plasmid (m) and MCP-5 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ccl12 homology arms to support homology-directed repair at defined Ccl12 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.