Mucin 5B/MUC5B Double Nickase Plasmid (h): sc-401077-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Mucin 5B/MUC5B 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
- Mucin 5B/MUC5B Double Nickase Plasmid (h) and Mucin 5B/MUC5B Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MUC5B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Mucin 5B/MUC5B Antibody (5B#19-2E): sc-21768
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Mucin 5B/MUC5B Double Nickase Plasmid (h) | sc-401077-NIC | 20 µg | $410.00 | |||
Mucin 5B/MUC5B Double Nickase Plasmid (h2) | sc-401077-NIC-2 | 20 µg | $410.00 |
MUC5B encodes mucin 5B, a high-molecular-weight, gel-forming secreted mucin that is heavily O-glycosylated and polymerizes through disulfide bonding to form the mucus barrier on airway and other mucosal epithelia. This barrier regulates mucociliary clearance, hydration, and innate defense by shaping the viscoelastic properties of mucus and influencing interactions with microbes and particulates. MUC5B expression is controlled by epithelial differentiation programs and inflammatory signaling pathways, including cytokine-driven transcriptional responses and ER–Golgi secretory processing that supports mucin biosynthesis. Dysregulated MUC5B levels or altered mucus properties have been implicated in chronic airway diseases and fibrotic lung pathology, making it a key target for studying epithelial homeostasis and mucosal defense mechanisms.
Mucin 5B/MUC5B Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MUC5B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MUC5B. 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 MUC5B 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 MUC5B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.