Lck Double Nickase Plasmid (h): sc-400434-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Lck 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
- Lck Double Nickase Plasmid (h) and Lck Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting LCK. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Lck Antibody (3A5): sc-433
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Lck Double Nickase Plasmid (h) | sc-400434-NIC | 20 µg | $410.00 | |||
Lck Double Nickase Plasmid (h2) | sc-400434-NIC-2 | 20 µg | $410.00 |
LCK encodes Lck, a Src family tyrosine kinase that initiates signaling downstream of the T cell receptor by phosphorylating ITAM motifs and coordinating recruitment of ZAP70 and adaptor complexes. Through these proximal events, Lck regulates activation of MAPK/ERK, PI3K–AKT, and NF-κB/NFAT pathways that control T cell development, activation, cytokine production, and immune synapse formation. Dysregulated LCK activity or expression is linked to altered lymphocyte signaling states and has been implicated in immune dysregulation and hematologic malignancy biology. As a node in receptor-proximal phosphorylation networks, Lck is widely studied in phosphoproteomics, kinase signaling, and functional genomics of human immune cells.
Lck Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the LCK locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within LCK. 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 LCK 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 LCK-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.