COL12A1 CRISPR/Cas9 KO Plasmid (h): sc-402361
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL12A1 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 COL12A1 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: COL12A1 Antibody (A-11): sc-166020
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL12A1 CRISPR/Cas9 KO Plasmid (h) | sc-402361 | 20 µg | $397.00 |
Overview
COL12A1 encodes collagen type XII alpha 1 chain, a fibril-associated collagen with interrupted triple helices (FACIT) that localizes to the extracellular matrix and associates with type I collagen fibrils. By linking fibrillar collagens to other matrix components, COL12A1 contributes to tissue tensile strength, mechanotransduction, and organization of the pericellular matrix, influencing cell adhesion and migration. Its function is tightly connected to extracellular matrix remodeling pathways and integrin-mediated signaling that shape stromal architecture during development and repair. Altered COL12A1 expression or variants have been implicated in heritable connective tissue phenotypes and musculoskeletal dysfunction, making it a useful target for studying matrix-driven disease mechanisms.
COL12A1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the COL12A1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the COL12A1 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 COL12A1 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 COL12A1 protein expression.
This CRISPR knockout system enables efficient generation of COL12A1-deficient cell models for investigation of COL12A1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting COL12A1 exon(s) critical for COL12A1 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 COL12A1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by COL12A1 CRISPR/Cas9 KO Plasmid (h) and COL12A1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the COL12A1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by COL12A1 HDR Plasmid (h) and COL12A1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by COL12A1 homology arms to support homology-directed repair at defined COL12A1 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.