TSSC3 CRISPR/Cas9 KO Plasmid (m): sc-423530
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TSSC3 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 TSSC3 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 | |
TSSC3 CRISPR/Cas9 KO Plasmid (m) | sc-423530 | 20 µg | $397.00 |
Overview
Phlda2 encodes the pleckstrin homology-like domain family A member 2 protein, also known as TSSC3, an imprinted, growth-regulatory factor implicated in controlling placental development and fetal growth in mouse. TSSC3 is linked to signaling processes that influence cellular proliferation and survival, including pathways integrating phosphoinositide-dependent cues and stress-responsive regulation of gene expression. Altered dosage or dysregulated imprinting at the Phlda2 locus has been associated with developmental phenotypes and has relevance to studies of growth control and tumor suppressor-like functions in specific cellular contexts. As a maternally expressed gene, Phlda2 is also a useful model for investigating parent-of-origin effects, epigenetic regulation, and dosage-sensitive transcriptional networks.
TSSC3 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Phlda2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Phlda2 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 Phlda2 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 TSSC3 protein expression.
This CRISPR knockout system enables efficient generation of Phlda2-deficient cell models for investigation of TSSC3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Phlda2 exon(s) critical for TSSC3 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 Phlda2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TSSC3 CRISPR/Cas9 KO Plasmid (m) and TSSC3 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Phlda2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TSSC3 HDR Plasmid (m) and TSSC3 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Phlda2 homology arms to support homology-directed repair at defined Phlda2 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.