Timeless Double Nickase Plasmid (h): sc-404555-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Timeless 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
- Timeless Double Nickase Plasmid (h) and Timeless Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TIMELESS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Timeless Antibody (A-3): sc-393122
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Timeless Double Nickase Plasmid (h) | sc-404555-NIC | 20 µg | $410.00 | |||
Timeless Double Nickase Plasmid (h2) | sc-404555-NIC-2 | 20 µg | $410.00 |
Human TIMELESS encodes Timeless, a conserved replication fork–associated factor that coordinates DNA replication with circadian and cell-cycle control. Timeless forms complexes with TIPIN and other replisome components to stabilize stalled forks, regulate the ATR–CHK1 checkpoint, and limit replication stress by promoting proper fork restart and S-phase progression. Through these functions it influences genome integrity, chromatin dynamics, and responses to DNA damage. Dysregulation of TIMELESS has been linked in the literature to altered checkpoint signaling and genomic instability phenotypes that are commonly interrogated in cancer biology and stress-response research models.
Timeless Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TIMELESS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TIMELESS. 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 TIMELESS 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 TIMELESS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.