TMEM65 CRISPR/Cas9 KO Plasmid (h): sc-406584
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TMEM65 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 TMEM65 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 | |
TMEM65 CRISPR/Cas9 KO Plasmid (h) | sc-406584 | 20 µg | $397.00 |
Overview
TMEM65 encodes transmembrane protein 65, a mitochondria-associated membrane protein implicated in maintaining inner mitochondrial membrane organization and supporting oxidative phosphorylation. TMEM65 has been linked to mitochondrial bioenergetics, regulation of membrane potential, and cellular responses to metabolic stress, thereby influencing ATP production and reactive oxygen species homeostasis. Altered TMEM65 function has been associated with mitochondrial dysfunction phenotypes and is relevant to studies of energy-demanding tissues where impaired respiration can contribute to neuromuscular and cardiometabolic disease mechanisms. As a mitochondrial pathway component, TMEM65 is useful for interrogating mitochondrial signaling, organelle quality control, and stress-adaptive transcriptional programs.
TMEM65 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TMEM65 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TMEM65 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 TMEM65 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 TMEM65 protein expression.
This CRISPR knockout system enables efficient generation of TMEM65-deficient cell models for investigation of TMEM65 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TMEM65 exon(s) critical for TMEM65 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 TMEM65 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TMEM65 CRISPR/Cas9 KO Plasmid (h) and TMEM65 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TMEM65 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TMEM65 HDR Plasmid (h) and TMEM65 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TMEM65 homology arms to support homology-directed repair at defined TMEM65 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.