DBT CRISPR/Cas9 KO Plasmid (h): sc-410755
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DBT 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 DBT 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DBT CRISPR/Cas9 KO Plasmid (h) | sc-410755 | 20 µg | $397.00 |
Overview
Dihydrolipoamide branched chain transacylase E2 (DBT) is the E2 core component of the mitochondrial branched-chain α-ketoacid dehydrogenase (BCKDH) complex, catalyzing acyl-transfer steps essential for branched-chain amino acid catabolism. DBT supports mitochondrial carbon flux and redox balance by enabling efficient oxidation of leucine, isoleucine, and valine-derived ketoacids, linking amino acid utilization to energy production. Its activity is coordinated with BCKDH regulatory phosphorylation and impacts metabolic stress responses, mitochondrial function, and nutrient-sensing pathways. Disruption or altered regulation of DBT and related BCKDH components is associated with inborn errors of metabolism affecting branched-chain amino acid homeostasis and is frequently studied in the context of metabolic rewiring and mitochondrial dysfunction.
DBT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the DBT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the DBT 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 DBT 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 DBT protein expression.
This CRISPR knockout system enables efficient generation of DBT-deficient cell models for investigation of DBT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting DBT exon(s) critical for DBT 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 DBT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by DBT CRISPR/Cas9 KO Plasmid (h) and DBT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the DBT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by DBT HDR Plasmid (h) and DBT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by DBT homology arms to support homology-directed repair at defined DBT 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.