E2A CRISPR/Cas9 KO Plasmid (m): sc-423303
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- E2A 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 E2A 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: E2A Antibody (Yae): sc-416
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
E2A CRISPR/Cas9 KO Plasmid (m) | sc-423303 | 20 µg | $397.00 |
Overview
Tcf3 encodes the E2A basic helix–loop–helix transcription factor, a key regulator of lineage commitment and differentiation programs, particularly within hematopoietic and lymphoid compartments. E2A forms dimers that bind E-box motifs to control gene networks involved in cell-cycle control, apoptosis, and developmental timing, and it functionally intersects with Notch, Wnt/β-catenin, and TGF-β signaling contexts. In mouse models, altered Tcf3 activity perturbs B- and T-cell development and can reshape transcriptional states that influence stem/progenitor maintenance. Dysregulation of E2A-dependent transcriptional circuitry is frequently studied in the context of oncogenic transcription factor networks and immune cell maturation defects.
E2A CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tcf3 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tcf3 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 Tcf3 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 E2A protein expression.
This CRISPR knockout system enables efficient generation of Tcf3-deficient cell models for investigation of E2A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tcf3 exon(s) critical for E2A 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 Tcf3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by E2A CRISPR/Cas9 KO Plasmid (m) and E2A CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tcf3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by E2A HDR Plasmid (m) and E2A HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tcf3 homology arms to support homology-directed repair at defined Tcf3 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.