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.
Myf-5 CRISPR/Cas9 KO Plasmid (h): sc-400449
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Myf-5 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 Myf-5 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: Myf-5 Antibody (B-2): sc-518039
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Myf-5 CRISPR/Cas9 KO Plasmid (h) | sc-400449 | 20 µg | $397.00 |
Overview
MYF5 encodes the human myogenic regulatory factor Myf-5, a basic helix-loop-helix transcription factor that functions early in skeletal muscle lineage determination and myoblast commitment. Myf-5 regulates gene expression programs that drive myogenic differentiation through heterodimerization with E-proteins and binding to E-box motifs in muscle-specific regulatory elements. It acts within developmental networks that coordinate myogenesis, including cross-regulation with other MRF family members such as MYOD1 and MYOG, influencing cell-fate decisions and differentiation timing. Dysregulation of MYF5-associated transcriptional control is relevant to studies of congenital myopathies, muscle regeneration defects, and tumor contexts showing aberrant myogenic factor expression or differentiation blockade.
Myf-5 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MYF5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the MYF5 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 MYF5 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 Myf-5 protein expression.
This CRISPR knockout system enables efficient generation of MYF5-deficient cell models for investigation of Myf-5 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting MYF5 exon(s) critical for Myf-5 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 MYF5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Myf-5 CRISPR/Cas9 KO Plasmid (h) and Myf-5 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the MYF5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Myf-5 HDR Plasmid (h) and Myf-5 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by MYF5 homology arms to support homology-directed repair at defined MYF5 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.