GLI-3 CRISPR/Cas9 KO Plasmid (m): sc-420577
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GLI-3 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 GLI-3 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: GLI-3 Antibody (B-4): sc-74478
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GLI-3 CRISPR/Cas9 KO Plasmid (m) | sc-420577 | 20 µg | $397.00 |
Overview
Gli3 encodes GLI-3, a zinc-finger transcription factor that functions as a context-dependent activator or repressor in Hedgehog signaling, integrating cues from primary cilia to regulate developmental gene expression programs. In mouse, GLI-3 is critical for patterning of the limb, neural tube, and craniofacial structures by controlling transcriptional outputs downstream of PTCH1/SMO and modulating GLI family activity. Proteolytic processing and post-translational regulation of GLI-3 help set morphogen gradients and boundary formation during embryogenesis. Dysregulated Gli3 activity is associated with congenital malformation phenotypes and is frequently used as a model to interrogate Hedgehog-dependent tissue homeostasis and oncogenic pathway rewiring.
GLI-3 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Gli3 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Gli3 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 Gli3 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 GLI-3 protein expression.
This CRISPR knockout system enables efficient generation of Gli3-deficient cell models for investigation of GLI-3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Gli3 exon(s) critical for GLI-3 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 Gli3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GLI-3 CRISPR/Cas9 KO Plasmid (m) and GLI-3 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Gli3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GLI-3 HDR Plasmid (m) and GLI-3 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Gli3 homology arms to support homology-directed repair at defined Gli3 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.