TRC8 CRISPR/Cas9 KO Plasmid (m): sc-429220
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TRC8 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 TRC8 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: TRC8 Antibody (H-9): sc-390347
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TRC8 CRISPR/Cas9 KO Plasmid (m) | sc-429220 | 20 µg | $397.00 |
Overview
Rnf139 encodes TRC8, an endoplasmic reticulum–associated, multi-pass membrane E3 ubiquitin ligase containing a RING finger domain. TRC8 contributes to ubiquitin-dependent control of protein stability, linking ER membrane quality control and sterol- and lipid-related regulation to broader proteostasis pathways. Through modulation of ubiquitination and turnover of select substrates, TRC8 can influence cellular growth control, stress responses, and signaling networks coordinated at the ER. Dysregulation of RNF139/TRC8-mediated ubiquitin signaling has been investigated in the context of altered proliferation and genome stability, making it relevant for mechanistic studies of disease-associated pathways in mouse models.
TRC8 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rnf139 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rnf139 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 Rnf139 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 TRC8 protein expression.
This CRISPR knockout system enables efficient generation of Rnf139-deficient cell models for investigation of TRC8 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rnf139 exon(s) critical for TRC8 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 Rnf139 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TRC8 CRISPR/Cas9 KO Plasmid (m) and TRC8 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rnf139 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TRC8 HDR Plasmid (m) and TRC8 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rnf139 homology arms to support homology-directed repair at defined Rnf139 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.