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.
UKHC Double Nickase Plasmid (h): sc-402349-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- UKHC 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
- UKHC Double Nickase Plasmid (h) and UKHC Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KIF5B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: UKHC Antibody (F-5): sc-133184
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
UKHC Double Nickase Plasmid (h) | sc-402349-NIC | 20 µg | $410.00 | |||
UKHC Double Nickase Plasmid (h2) | sc-402349-NIC-2 | 20 µg | $410.00 |
KIF5B encodes the ubiquitously expressed kinesin-1 heavy chain (UKHC), a microtubule-dependent motor that powers ATP-driven anterograde transport of membranous cargo, organelles, and protein complexes along axons and other polarized cell types. Through interactions with kinesin light chains and cargo adaptors, KIF5B coordinates intracellular trafficking, contributes to mitotic spindle dynamics, and supports maintenance of cell polarity and cytoskeletal organization. These functions intersect with pathways governing neuronal transport, mitochondrial distribution, and vesicle trafficking in secretory and endocytic systems. Altered KIF5B regulation or rearrangements have been linked to oncogenic signaling contexts and disrupted trafficking phenotypes relevant to tumor biology and neurobiology.
UKHC Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KIF5B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KIF5B. 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 KIF5B 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 KIF5B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.