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.
smoothelin CRISPR/Cas9 KO Plasmid (h): sc-400923
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- smoothelin 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 smoothelin 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: smoothelin Antibody (C-8): sc-376902
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
smoothelin CRISPR/Cas9 KO Plasmid (h) | sc-400923 | 20 µg | $397.00 |
Overview
SMTN encodes smoothelin, a cytoskeletal protein selectively enriched in fully differentiated smooth muscle cells where it associates with actin filament organization and contractile apparatus integrity. Smoothelin contributes to regulation of smooth muscle contractility, cellular architecture, and phenotypic stability during vascular and visceral smooth muscle remodeling. Altered SMTN expression patterns are used as markers of smooth muscle differentiation state and are implicated in processes such as vascular remodeling, fibrosis-associated myofibroblast transitions, and changes in vessel tone. As a lineage-restricted cytoskeletal component, SMTN is relevant for studying mechanisms that govern smooth muscle maturation, mechanotransduction, and cytoskeletal signaling networks.
smoothelin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SMTN gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SMTN 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 SMTN 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 smoothelin protein expression.
This CRISPR knockout system enables efficient generation of SMTN-deficient cell models for investigation of smoothelin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SMTN exon(s) critical for smoothelin 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 SMTN genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by smoothelin CRISPR/Cas9 KO Plasmid (h) and smoothelin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SMTN locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by smoothelin HDR Plasmid (h) and smoothelin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SMTN homology arms to support homology-directed repair at defined SMTN 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.