Myf-5 Double Nickase Plasmid (h): sc-400449-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Myf-5 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Myf-5 Double Nickase Plasmid (h) and Myf-5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MYF5. One or both designs may be available
- 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 Double Nickase Plasmid (h) | sc-400449-NIC | 20 µg | $410.00 | |||
Myf-5 Double Nickase Plasmid (h2) | sc-400449-NIC-2 | 20 µg | $410.00 |
MYF5 encodes the basic helix–loop–helix transcription factor Myf-5, an early myogenic regulatory factor that helps specify skeletal muscle lineage and initiate myoblast determination. Myf-5 functions through E-box DNA binding and cooperative transcriptional control with other MRFs and cofactors to regulate genes involved in myogenesis, cell-cycle exit, and differentiation. Its activity intersects with developmental signaling pathways such as WNT, SHH, and NOTCH that shape muscle progenitor fate decisions. Dysregulation of MYF5 expression or myogenic programs is relevant to studies of congenital myopathies, muscle wasting and regeneration, and tumor contexts where myogenic lineage markers are aberrantly expressed.
Myf-5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MYF5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MYF5. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt MYF5 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of MYF5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.