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.
Twinkle CRISPR/Cas9 KO Plasmid (h): sc-404159
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Twinkle 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 Twinkle 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: Twinkle Antibody (1C5): sc-293368
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Twinkle CRISPR/Cas9 KO Plasmid (h) | sc-404159 | 20 µg | $397.00 |
Overview
TWNK encodes Twinkle, a mitochondrial DNA helicase essential for mtDNA replication and maintenance of nucleoids, functioning with the mitochondrial replisome to support oxidative phosphorylation capacity. By unwinding duplex mtDNA during replication, Twinkle helps preserve mtDNA copy number and genome integrity, influencing mitochondrial transcription, respiratory chain biogenesis, and cellular energy homeostasis. Perturbation of TWNK is linked to mitochondrial genome instability, including mtDNA depletion or multiple deletions, and is associated with neuromuscular and neurodegenerative mitochondrial disease phenotypes. TWNK loss-of-function studies are therefore relevant for investigating mitochondrial stress signaling, bioenergetic remodeling, and replication-associated mtDNA damage responses.
Twinkle CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TWNK gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TWNK 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 TWNK 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 Twinkle protein expression.
This CRISPR knockout system enables efficient generation of TWNK-deficient cell models for investigation of Twinkle signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TWNK exon(s) critical for Twinkle 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 TWNK genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Twinkle CRISPR/Cas9 KO Plasmid (h) and Twinkle CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TWNK locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Twinkle HDR Plasmid (h) and Twinkle HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TWNK homology arms to support homology-directed repair at defined TWNK 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.