EN2/Engrailed 2 CRISPR/Cas9 KO Plasmid (m): sc-420172
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EN2/Engrailed 2 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 EN2/Engrailed 2 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 | |
EN2/Engrailed 2 CRISPR/Cas9 KO Plasmid (m) | sc-420172 | 20 µg | $397.00 |
Overview
Engrailed 2 (EN2) is a homeobox transcription factor that helps pattern the developing midbrain–hindbrain region and regulates neuronal differentiation, axon guidance, and regional identity programs in the mouse central nervous system. By binding DNA through its homeodomain, EN2 coordinates transcriptional networks downstream of morphogen gradients such as FGF and WNT signaling that shape cerebellar and tectal development. Altered EN2 dosage or regulatory control has been linked to neurodevelopmental phenotypes and behavioral traits, making it a useful node for studying gene regulatory circuitry in the brain. EN2 is also used as a marker and mechanistic entry point for investigating transcriptional control of neural progenitor fate and circuit formation.
EN2/Engrailed 2 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the En2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the En2 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 En2 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 EN2/Engrailed 2 protein expression.
This CRISPR knockout system enables efficient generation of En2-deficient cell models for investigation of EN2/Engrailed 2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting En2 exon(s) critical for EN2/Engrailed 2 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 En2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by EN2/Engrailed 2 CRISPR/Cas9 KO Plasmid (m) and EN2/Engrailed 2 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the En2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by EN2/Engrailed 2 HDR Plasmid (m) and EN2/Engrailed 2 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by En2 homology arms to support homology-directed repair at defined En2 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.