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.
ACTA2 CRISPR/Cas9 KO Plasmid (h): sc-400003
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ACTA2 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 ACTA2 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: α-Actin Antibody (1A4): sc-32251
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ACTA2 CRISPR/Cas9 KO Plasmid (h) | sc-400003 | 20 µg | $397.00 |
Overview
ACTA2 encodes alpha-smooth muscle actin (α-SMA), a core component of the contractile apparatus in vascular smooth muscle cells and activated myofibroblasts. ACTA2 integrates into actin stress fibers to regulate cell contractility, cytoskeletal organization, and mechanotransduction, influencing processes such as migration, adhesion, and extracellular matrix remodeling. Through actin dynamics and RhoA/ROCK-dependent contractile signaling, ACTA2 contributes to vascular tone and tissue stiffness responses. Dysregulated ACTA2 expression or function is linked to vascular pathobiology and fibroproliferative remodeling, supporting its relevance in studies of smooth muscle differentiation and fibrosis-associated phenotypes.
ACTA2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ACTA2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ACTA2 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 ACTA2 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 ACTA2 protein expression.
This CRISPR knockout system enables efficient generation of ACTA2-deficient cell models for investigation of ACTA2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ACTA2 exon(s) critical for ACTA2 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 ACTA2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ACTA2 CRISPR/Cas9 KO Plasmid (h) and ACTA2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ACTA2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ACTA2 HDR Plasmid (h) and ACTA2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ACTA2 homology arms to support homology-directed repair at defined ACTA2 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.