Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Evi-1 CRISPR/Cas9 KO Plasmid (h): sc-404112
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Evi-1 CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 Evi-1 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: Evi-1 Antibody (H-8): sc-515456
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Evi-1 CRISPR/Cas9 KO Plasmid (h) | sc-404112 | 20 µg | $397.00 |
Overview
MECOM encodes the transcription factor Evi-1, a zinc finger DNA-binding protein that regulates gene expression programs controlling hematopoietic stem and progenitor cell maintenance, lineage commitment, and cellular differentiation. Evi-1 participates in transcriptional and epigenetic networks that intersect with TGF-β/SMAD signaling, PI3K–AKT pathways, and chromatin remodeling to modulate proliferation and survival. Dysregulated MECOM/Evi-1 activity is frequently associated with hematologic malignancies, where altered transcriptional control contributes to aberrant self-renewal and impaired differentiation. In addition to blood development, MECOM has been implicated in developmental processes and oncogenic transcriptional circuitry relevant to transformation and metastatic phenotypes in multiple tissue contexts.
Evi-1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MECOM gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the MECOM 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 MECOM 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 Evi-1 protein expression.
This CRISPR knockout system enables efficient generation of MECOM-deficient cell models for investigation of Evi-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting MECOM exon(s) critical for Evi-1 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 MECOM genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Evi-1 CRISPR/Cas9 KO Plasmid (h) and Evi-1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the MECOM locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Evi-1 HDR Plasmid (h) and Evi-1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by MECOM homology arms to support homology-directed repair at defined MECOM 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.