spectrin α I CRISPR/Cas9 KO Plasmid (h): sc-403563
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- spectrin α I 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 spectrin α I 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: spectrin α I Antibody (B-12): sc-271130
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
spectrin α I CRISPR/Cas9 KO Plasmid (h) | sc-403563 | 20 µg | $397.00 |
Overview
SPTA1 encodes human spectrin α I, a core component of the erythrocyte membrane skeleton that forms heterodimers with β-spectrin to assemble a lattice supporting membrane stability and deformability. This cytoskeletal network links integral membrane proteins to the actin-based junctional complex, coordinating mechanical resilience during repeated cycles of shear stress in circulation. Disruption of spectrin architecture perturbs red cell shape maintenance and membrane integrity, influencing processes such as membrane trafficking, cytoskeletal remodeling, and ion homeostasis. Genetic variation or deficiency in SPTA1 is associated with inherited red blood cell membrane disorders, including hereditary spherocytosis and elliptocytosis, making it a relevant target for studying genotype–phenotype relationships in erythroid biology.
spectrin α I CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SPTA1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SPTA1 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 SPTA1 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 spectrin α I protein expression.
This CRISPR knockout system enables efficient generation of SPTA1-deficient cell models for investigation of spectrin α I signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SPTA1 exon(s) critical for spectrin α I 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 SPTA1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by spectrin α I CRISPR/Cas9 KO Plasmid (h) and spectrin α I CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SPTA1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by spectrin α I HDR Plasmid (h) and spectrin α I HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SPTA1 homology arms to support homology-directed repair at defined SPTA1 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.