TMPIT CRISPR/Cas9 KO Plasmid (m): sc-431998
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TMPIT 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 TMPIT 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 | |
TMPIT CRISPR/Cas9 KO Plasmid (m) | sc-431998 | 20 µg | $397.00 |
Overview
Tmem120a encodes TMPIT, a multi-pass membrane protein predominantly associated with the nuclear envelope and endoplasmic reticulum, where it has been linked to lipid metabolism and cellular mechanotransduction programs. Studies implicate TMPIT in adipocyte differentiation and regulation of lipid storage, connecting Tmem120a to pathways controlling membrane lipid composition and metabolic homeostasis. Altered TMPIT function has been explored in the context of adipose tissue biology, obesity-associated phenotypes, and peripheral sensory mechanisms related to mechanical stimuli. In mouse model systems, Tmem120a perturbation provides a route to interrogate how membrane-associated proteins shape gene expression and organelle function in metabolically active tissues.
TMPIT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tmem120a gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tmem120a 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 Tmem120a 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 TMPIT protein expression.
This CRISPR knockout system enables efficient generation of Tmem120a-deficient cell models for investigation of TMPIT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tmem120a exon(s) critical for TMPIT 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 Tmem120a genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TMPIT CRISPR/Cas9 KO Plasmid (m) and TMPIT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tmem120a locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TMPIT HDR Plasmid (m) and TMPIT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tmem120a homology arms to support homology-directed repair at defined Tmem120a 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.