SAP 49 Double Nickase Plasmid (h): sc-407057-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SAP 49 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
- SAP 49 Double Nickase Plasmid (h) and SAP 49 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SF3B4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: SAP 49 Antibody (G-12): sc-365570
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SAP 49 Double Nickase Plasmid (h) | sc-407057-NIC | 20 µg | $410.00 | |||
SAP 49 Double Nickase Plasmid (h2) | sc-407057-NIC-2 | 20 µg | $410.00 |
SF3B4 encodes SAP 49, a core component of the SF3b subcomplex within the U2 small nuclear ribonucleoprotein that helps recognize branch point sequences during pre-mRNA splicing. By supporting spliceosome assembly and splice site selection, SAP 49 contributes to transcriptome integrity and downstream regulation of RNA processing, cell-cycle programs, and stress responses. Perturbation of SF3B4-dependent splicing is linked to altered isoform expression and broad changes in gene regulatory networks observed in multiple disease contexts, including cancers and developmental disorders. As a spliceosomal factor, SAP 49 is frequently studied in mechanisms of splicing fidelity, alternative splicing control, and genotype-to-isoform relationships.
SAP 49 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SF3B4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SF3B4. 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 SF3B4 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 SF3B4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.