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.
mSHMT Double Nickase Plasmid (h): sc-402609-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- mSHMT 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
- mSHMT Double Nickase Plasmid (h) and mSHMT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SHMT2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: mSHMT Antibody (F-11): sc-390641
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
mSHMT Double Nickase Plasmid (h) | sc-402609-NIC | 20 µg | $410.00 | |||
mSHMT Double Nickase Plasmid (h2) | sc-402609-NIC-2 | 20 µg | $410.00 |
SHMT2 encodes mitochondrial serine hydroxymethyltransferase (mSHMT), a PLP-dependent enzyme that catalyzes the reversible conversion of serine to glycine while generating 5,10-methylenetetrahydrofolate in the mitochondrial one-carbon cycle. This activity supports folate-mediated one-carbon metabolism, coupling amino acid interconversion to nucleotide biosynthesis, redox homeostasis, and mitochondrial function. Through its role in sustaining formate supply to the cytosolic folate pool, SHMT2 influences DNA replication and repair capacity as well as cellular responses to metabolic and oxidative stress. Dysregulation of SHMT2-linked pathways has been associated with proliferative and metabolic phenotypes and is studied in contexts including tumor metabolism and mitochondrial dysfunction.
mSHMT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SHMT2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SHMT2. 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 SHMT2 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 SHMT2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.