PIMT CRISPR/Cas9 KO Plasmid (h): sc-407101
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PIMT 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 PIMT 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 | |
PIMT CRISPR/Cas9 KO Plasmid (h) | sc-407101 | 20 µg | $397.00 |
Overview
Human TGS1 encodes trimethylguanosine synthase 1 (PIMT), a SAM-dependent methyltransferase that converts the 2,2,7-trimethylguanosine (TMG) cap on snRNAs and other small nuclear RNAs. This modification supports snRNP maturation, nuclear import, and spliceosome function, linking TGS1 activity to pre-mRNA splicing fidelity and broader RNA processing networks. TGS1 also interfaces with RNA quality control and ribonucleoprotein biogenesis pathways that influence proteostasis and stress responses. Dysregulation of RNA metabolism and splicing-associated programs implicates TGS1/PIMT in mechanisms relevant to neurodegeneration, cancer biology, and developmental phenotypes, making it a useful node for pathway-focused studies.
PIMT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TGS1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TGS1 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 TGS1 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 PIMT protein expression.
This CRISPR knockout system enables efficient generation of TGS1-deficient cell models for investigation of PIMT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TGS1 exon(s) critical for PIMT 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 TGS1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PIMT CRISPR/Cas9 KO Plasmid (h) and PIMT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TGS1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PIMT HDR Plasmid (h) and PIMT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TGS1 homology arms to support homology-directed repair at defined TGS1 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.