DDT CRISPR/Cas9 KO Plasmid (h): sc-418461
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DDT CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the DDT genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DDT Antibody (1G1): sc-517061
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DDT CRISPR/Cas9 KO Plasmid (h) | sc-418461 | 20 µg | $397.00 |
Overview
DDT (D-dopachrome tautomerase), also known as MIF-2, is a cytokine-like enzyme that participates in immune signaling and cellular stress responses, functioning alongside the MIF axis. It contributes to regulation of inflammatory mediator production, leukocyte recruitment, and pro-survival signaling through pathways that intersect with MAPK/ERK and NF-κB networks, depending on cellular context and receptor usage. DDT expression has been linked to modulation of innate and adaptive immune activity, with reported relevance to tumor-associated inflammation, autoimmune phenotypes, and metabolic dysregulation. As a soluble factor with enzymatic activity, DDT provides a tractable node for dissecting how cytokine signaling integrates with redox, proliferation, and migration programs in human cells.
DDT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the DDT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the DDT together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the DDT open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish DDT protein expression.
This CRISPR knockout system enables efficient generation of DDT-deficient cell models for investigation of DDT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting DDT exon(s) critical for DDT function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple DDT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by DDT CRISPR/Cas9 KO Plasmid (h) and DDT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the DDT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by DDT HDR Plasmid (h) and DDT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by DDT homology arms to support homology-directed repair at defined DDT target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.