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.
NIRF CRISPR/Cas9 KO Plasmid (h): sc-403908
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NIRF 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 NIRF 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: NIRF Antibody (C-10): sc-398953
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NIRF CRISPR/Cas9 KO Plasmid (h) | sc-403908 | 20 µg | $397.00 |
Overview
UHRF2 encodes NIRF, a RING finger E3 ubiquitin ligase and epigenetic regulator implicated in coupling ubiquitin signaling to chromatin-based control of gene expression. NIRF contains domains that support recognition of modified histones and DNA methylation states, linking it to maintenance of epigenetic information during DNA replication and to regulation of cell-cycle progression. Through ubiquitin-mediated proteostasis and chromatin remodeling, UHRF2 can influence DNA damage responses and transcriptional programs that govern proliferation and differentiation. Dysregulation of UHRF2/NIRF expression or activity has been associated with altered epigenetic landscapes observed in multiple cancer and neurobiology contexts, making it relevant for studies of genome stability and oncogenic signaling.
NIRF CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the UHRF2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the UHRF2 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 UHRF2 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 NIRF protein expression.
This CRISPR knockout system enables efficient generation of UHRF2-deficient cell models for investigation of NIRF signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting UHRF2 exon(s) critical for NIRF 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 UHRF2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by NIRF CRISPR/Cas9 KO Plasmid (h) and NIRF CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the UHRF2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by NIRF HDR Plasmid (h) and NIRF HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by UHRF2 homology arms to support homology-directed repair at defined UHRF2 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.