RNF185 CRISPR/Cas9 KO Plasmid (h): sc-416413
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNF185 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 RNF185 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 | |
RNF185 CRISPR/Cas9 KO Plasmid (h) | sc-416413 | 20 µg | $397.00 |
Overview
RNF185 encodes a mitochondrial outer membrane RING-type E3 ubiquitin ligase that contributes to protein quality control by catalyzing ubiquitin transfer to selected substrates. Through ubiquitin-dependent turnover and signaling, RNF185 has been linked to regulation of mitochondrial homeostasis, mitophagy-related processes, and crosstalk between mitochondrial function and innate immune signaling. Reported interactions with autophagy and inflammatory pathways support its relevance for studying stress responses that couple organelle damage to downstream transcriptional programs. Altered RNF185 activity has been investigated in contexts where mitochondrial dysfunction and dysregulated proteostasis are implicated, including neuroinflammation and cancer-associated metabolic remodeling.
RNF185 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RNF185 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RNF185 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 RNF185 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 RNF185 protein expression.
This CRISPR knockout system enables efficient generation of RNF185-deficient cell models for investigation of RNF185 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RNF185 exon(s) critical for RNF185 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 RNF185 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RNF185 CRISPR/Cas9 KO Plasmid (h) and RNF185 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RNF185 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RNF185 HDR Plasmid (h) and RNF185 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RNF185 homology arms to support homology-directed repair at defined RNF185 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.