Dorfin CRISPR/Cas9 KO Plasmid (h): sc-406860
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Dorfin 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 Dorfin 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 | |
Dorfin CRISPR/Cas9 KO Plasmid (h) | sc-406860 | 20 µg | $397.00 |
Overview
RNF19A encodes Dorfin, an RBR family E3 ubiquitin ligase implicated in ubiquitin-dependent protein quality control and proteostasis. Dorfin participates in the recognition and ubiquitination of misfolded or aggregation-prone substrates, linking endoplasmic reticulum stress responses and cytosolic degradation pathways to maintain cellular homeostasis. Through regulation of ubiquitin signaling and turnover of neuronal and stress-associated proteins, RNF19A has been studied in the context of neurodegeneration and other conditions characterized by protein aggregation and dysregulated proteostasis. Its activity intersects with pathways governing ER-associated degradation, autophagy-related clearance, and stress-induced signaling networks.
Dorfin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the RNF19A gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the RNF19A 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 RNF19A 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 Dorfin protein expression.
This CRISPR knockout system enables efficient generation of RNF19A-deficient cell models for investigation of Dorfin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting RNF19A exon(s) critical for Dorfin 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 RNF19A genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Dorfin CRISPR/Cas9 KO Plasmid (h) and Dorfin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the RNF19A locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Dorfin HDR Plasmid (h) and Dorfin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by RNF19A homology arms to support homology-directed repair at defined RNF19A 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.