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.
HRF CRISPR/Cas9 KO Plasmid (m): sc-423518
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HRF 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 HRF 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: HRF Antibody (B-3): sc-133131
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HRF CRISPR/Cas9 KO Plasmid (m) | sc-423518 | 20 µg | $397.00 |
Overview
Mouse Tpt1 encodes histamine-releasing factor (HRF), also known as translationally controlled tumor protein, a conserved cytosolic protein implicated in regulation of cell growth, stress responses, and survival signaling. HRF participates in pathways linked to calcium homeostasis, apoptosis control, and modulation of inflammatory and immune-cell functions, with reported roles in cytokine release and cellular activation programs. Altered HRF/TPT1 activity has been associated with dysregulated inflammation and immune signaling as well as abnormal proliferative phenotypes relevant to oncology and tissue remodeling. These features make Tpt1 a useful target for dissecting stress-adaptation mechanisms, immune regulation, and cell-fate decisions in mouse model systems.
HRF CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tpt1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tpt1 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 Tpt1 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 HRF protein expression.
This CRISPR knockout system enables efficient generation of Tpt1-deficient cell models for investigation of HRF signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tpt1 exon(s) critical for HRF 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 Tpt1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by HRF CRISPR/Cas9 KO Plasmid (m) and HRF CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tpt1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by HRF HDR Plasmid (m) and HRF HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tpt1 homology arms to support homology-directed repair at defined Tpt1 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.