GAK Double Nickase Plasmid (h): sc-404235-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GAK 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
- GAK Double Nickase Plasmid (h) and GAK Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GAK. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GAK Antibody (D-2): sc-137053
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GAK Double Nickase Plasmid (h) | sc-404235-NIC | 20 µg | $410.00 | |||
GAK Double Nickase Plasmid (h2) | sc-404235-NIC-2 | 20 µg | $410.00 |
Cyclin G–associated kinase (GAK) is a serine/threonine kinase that couples clathrin-mediated membrane trafficking to cell-cycle control by regulating adaptor proteins and coat dynamics during endocytosis. In human cells, GAK supports receptor internalization and endosomal sorting, influencing signaling pathways such as EGFR trafficking and downstream mitogenic responses. It also contributes to Golgi and centrosome-associated processes linked to mitotic progression and proteostasis. Dysregulated GAK activity or expression has been associated with proliferative phenotypes and has been studied in the context of neurodegeneration-related pathways involving vesicle transport and protein turnover.
GAK Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GAK locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GAK. 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 GAK 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 GAK-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.