GSC CRISPR/Cas9 KO Plasmid (m): sc-420701
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GSC 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 GSC 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: GSC Antibody (L-36): sc-81964
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GSC CRISPR/Cas9 KO Plasmid (m) | sc-420701 | 20 µg | $397.00 |
Overview
Mouse Gsc encodes the homeobox transcription factor GSC, a conserved regulator of embryonic patterning and tissue morphogenesis. GSC functions downstream of major developmental signaling inputs, including TGF-β/BMP and Wnt pathways, to coordinate gene expression programs that control gastrulation movements, axial specification, and craniofacial development. Altered regulation of homeobox transcriptional networks is broadly relevant to congenital malformations and to contexts where developmental programs are reactivated in disease models. Accordingly, Gsc is frequently studied in transcriptional control, lineage commitment, and gene regulatory circuit analyses in mouse systems.
GSC CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Gsc gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Gsc 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 Gsc 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 GSC protein expression.
This CRISPR knockout system enables efficient generation of Gsc-deficient cell models for investigation of GSC signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Gsc exon(s) critical for GSC 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 Gsc genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GSC CRISPR/Cas9 KO Plasmid (m) and GSC CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Gsc locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GSC HDR Plasmid (m) and GSC HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Gsc homology arms to support homology-directed repair at defined Gsc 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.