SEMA5A CRISPR/Cas9 KO Plasmid (m): sc-422885
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SEMA5A 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 SEMA5A 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SEMA5A CRISPR/Cas9 KO Plasmid (m) | sc-422885 | 20 µg | $397.00 |
Overview
Sema5a encodes the transmembrane guidance cue SEMA5A, a member of the semaphorin family that regulates axon pathfinding, neuronal migration, and synapse formation through receptor-mediated signaling that remodels the cytoskeleton. In addition to roles in the nervous system, SEMA5A contributes to cell adhesion and motility programs in multiple tissues, intersecting with pathways that shape directional movement and tissue patterning. Altered SEMA5A expression or signaling has been associated with neurodevelopmental phenotypes and has been studied in contexts involving aberrant connectivity and inflammatory microenvironments. In mouse models, Sema5a perturbation is used to dissect mechanisms of circuit assembly and to link guidance signaling with cell state changes during development and disease-relevant processes.
SEMA5A CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Sema5a gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Sema5a 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 Sema5a 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 SEMA5A protein expression.
This CRISPR knockout system enables efficient generation of Sema5a-deficient cell models for investigation of SEMA5A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Sema5a exon(s) critical for SEMA5A 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 Sema5a genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SEMA5A CRISPR/Cas9 KO Plasmid (m) and SEMA5A CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Sema5a locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SEMA5A HDR Plasmid (m) and SEMA5A HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Sema5a homology arms to support homology-directed repair at defined Sema5a 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.