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.
MITF CRISPR/Cas9 KO Plasmid (m2): sc-421654-KO-2
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MITF CRISPR/Cas9 Knockout (KO) Plasmid (m2) 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 MITF 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: MITF Antibody (D-9): sc-515925
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MITF CRISPR/Cas9 KO Plasmid (m2) | sc-421654-KO-2 | 20 µg | $397.00 |
Overview
Mitf encodes microphthalmia-associated transcription factor (MITF), a basic helix-loop-helix leucine zipper regulator that coordinates lineage specification and differentiation programs in melanocytes and related neural crest–derived cell types. MITF controls transcriptional networks governing pigmentation, melanosome biogenesis, cell-cycle progression, and survival, integrating signaling inputs such as MAPK/ERK and cAMP/CREB pathways. In mouse models, altered Mitf activity impacts melanocyte development, pigmentary phenotypes, and immune-related functions in mast cells, linking the gene to mechanistic studies of melanogenesis and lineage plasticity. Dysregulation of MITF-dependent programs is widely used to interrogate pathways relevant to melanoma biology, including differentiation state switching and stress responses.
MITF CRISPR/Cas9 KO Plasmid (m2) is a pool of plasmids designed for targeted disruption of the Mitf gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Mitf 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 Mitf 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 MITF protein expression.
This CRISPR knockout system enables efficient generation of Mitf-deficient cell models for investigation of MITF signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Mitf exon(s) critical for MITF 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 Mitf genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MITF CRISPR/Cas9 KO Plasmid (m) and MITF CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Mitf locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MITF HDR Plasmid (m) and MITF HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Mitf homology arms to support homology-directed repair at defined Mitf 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.