SEMA5B CRISPR/Cas9 KO Plasmid (m): sc-422886
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SEMA5B 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 SEMA5B 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 | |
SEMA5B CRISPR/Cas9 KO Plasmid (m) | sc-422886 | 20 µg | $397.00 |
Overview
Sema5b encodes the transmembrane guidance cue SEMA5B, a member of the semaphorin family that regulates contact-dependent signaling during neural development and synaptic circuit assembly. SEMA5B influences axon guidance, dendritic patterning, and neurite outgrowth through pathways involving plexin receptors and downstream cytoskeletal remodeling. In mouse systems, altered Sema5b activity has been linked to changes in neuronal connectivity and plasticity, supporting its relevance to studies of neurodevelopmental and neuropsychiatric disease mechanisms. Its expression in the nervous system also makes it a useful node for investigating cell–cell communication programs that shape tissue patterning.
SEMA5B CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Sema5b gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Sema5b 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 Sema5b 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 SEMA5B protein expression.
This CRISPR knockout system enables efficient generation of Sema5b-deficient cell models for investigation of SEMA5B signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Sema5b exon(s) critical for SEMA5B 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 Sema5b genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SEMA5B CRISPR/Cas9 KO Plasmid (m) and SEMA5B CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Sema5b locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SEMA5B HDR Plasmid (m) and SEMA5B HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Sema5b homology arms to support homology-directed repair at defined Sema5b 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.