MIA CRISPR/Cas9 KO Plasmid (h): sc-404978
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MIA 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 MIA 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: MIA Antibody (C-10): sc-377375
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MIA CRISPR/Cas9 KO Plasmid (h) | sc-404978 | 20 µg | $397.00 |
Overview
Human MIA (melanoma inhibitory activity) encodes a secreted extracellular matrix–interacting protein implicated in regulation of cell adhesion, migration, and tissue remodeling. MIA can modulate integrin- and matrix-dependent signaling and influence cytoskeletal dynamics that shape invasive phenotypes and cellular stress responses. Altered MIA expression has been reported in melanoma and other malignancies, where it is frequently studied as a context-dependent mediator of tumor–microenvironment interactions and metastatic behavior. As a soluble factor, MIA is also used to interrogate paracrine signaling and extracellular pathway dependencies in cell-based models.
MIA CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MIA gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the MIA 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 MIA 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 MIA protein expression.
This CRISPR knockout system enables efficient generation of MIA-deficient cell models for investigation of MIA signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting MIA exon(s) critical for MIA 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 MIA genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MIA CRISPR/Cas9 KO Plasmid (h) and MIA CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the MIA locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MIA HDR Plasmid (h) and MIA HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by MIA homology arms to support homology-directed repair at defined MIA 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.