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.
LIS1 Double Nickase Plasmid (m): sc-422102-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- LIS1 Double Nickase Plasmid (m) 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
- LIS1 Double Nickase Plasmid (m) and LIS1 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Pafah1b1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: LIS1 Antibody (H-7): sc-374586
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
LIS1 Double Nickase Plasmid (m) | sc-422102-NIC | 20 µg | $410.00 | |||
LIS1 Double Nickase Plasmid (m2) | sc-422102-NIC-2 | 20 µg | $410.00 |
Mouse Pafah1b1 encodes LIS1, a dynein regulatory protein essential for microtubule-based transport, mitotic spindle orientation, and coupling of the dynein–dynactin motor to cargo during neuronal migration. LIS1 functions with NDE1/NDEL1 and cytoplasmic dynein to control centrosome positioning, nuclear translocation, and cortical development, linking it to pathways that govern cell polarity and division. Disruption of LIS1-dependent processes is strongly associated with neurodevelopmental phenotypes, including lissencephaly-related defects in neuronal layering and axon guidance. As a result, Pafah1b1 is widely studied in models of brain development, cilia- and microtubule-associated dynamics, and dynein-driven trafficking.
LIS1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Pafah1b1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Pafah1b1. 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 Pafah1b1 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 Pafah1b1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.