ICMT CRISPR/Cas9 KO Plasmid (h): sc-406029
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ICMT 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 ICMT 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 | |
ICMT CRISPR/Cas9 KO Plasmid (h) | sc-406029 | 20 µg | $397.00 |
Overview
ICMT (isoprenylcysteine carboxyl methyltransferase) is an endoplasmic reticulum–localized enzyme that catalyzes methyl esterification of prenylated cysteine residues at the C-terminus of CAAX proteins following proteolysis, a terminal step in protein prenylation. This modification tunes membrane association and subcellular trafficking of small GTPases and related signaling proteins, influencing pathways such as RAS/MAPK, PI3K–AKT, and cytoskeletal regulation. By regulating localization and activity of prenylated substrates, ICMT contributes to control of cell growth, polarity, and vesicular transport. Altered prenylation dynamics and ICMT-dependent processing have been linked to oncogenic signaling and other disorders where mislocalized prenylated proteins disrupt cellular homeostasis.
ICMT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ICMT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ICMT 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 ICMT 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 ICMT protein expression.
This CRISPR knockout system enables efficient generation of ICMT-deficient cell models for investigation of ICMT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ICMT exon(s) critical for ICMT 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 ICMT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ICMT CRISPR/Cas9 KO Plasmid (h) and ICMT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ICMT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ICMT HDR Plasmid (h) and ICMT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ICMT homology arms to support homology-directed repair at defined ICMT 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.