DNA pol θ Double Nickase Plasmid (h): sc-407414-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA pol θ 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
- DNA pol θ Double Nickase Plasmid (h) and DNA pol θ Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting POLQ. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA pol θ Double Nickase Plasmid (h) | sc-407414-NIC | 20 µg | $410.00 |
POLQ encodes DNA polymerase theta (Pol θ), an error-prone A-family polymerase that also possesses helicase-like activity and functions as a central mediator of microhomology-mediated end joining (MMEJ), also referred to as theta-mediated end joining. Pol θ operates at DNA double-strand breaks and stalled replication forks, promoting end joining using short microhomologies and thereby influencing genome stability, replication stress tolerance, and mutational signatures. This pathway interfaces with classical non-homologous end joining and homologous recombination, shaping repair pathway choice under conditions of compromised DNA end resection or HR capacity. Altered POLQ activity and expression are frequently studied in the context of chromosomal rearrangements and hypermutagenesis associated with cancer biology and DNA repair–defective states.
DNA pol θ Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the POLQ locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within POLQ. 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 POLQ 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 POLQ-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.